Regulatory DNA sequences of the human catalytic telomerase sub-unit gene, diagnostic and therapeutic use thereof

ABSTRACT

This invention relates to regulatory DNA sequences, comprising promoter sequences and intron sequences, for the gene for the human catalytic telomerase subunit. In addition, this invention relates to the use of these DNA sequences for pharmaceutical, diagnostic and therapeutic purposes, especially in the treatment of cancer and ageing.

STRUCTURE AND FUNCTION OF THE CHROMOSOME ENDS

The genetic material of eukaryotic cells is distributed on linear chromosomes. The ends of hereditary units are termed telomeres, derived from the Greek words telos (end) and meros (part, segment). Most telomeres consist of repeats of short sequences which are mainly composed of thymine and guanine (Zakian, 1995). In all the vertebrates which have so far been investigated, the telomeres consist of the sequence TTAGGG (Meyne et al., 1989).

The telomeres have a variety of important functions. They prevent the fusion of chromosomes (McClintock, 1941) and thus the formation of dicentric hereditary units. Such chromosomes having two centromeres can lead to the development of cancer due to loss of heterozygosis or duplication, or loss of genes.

In addition, telomeres serve the purpose of distinguishing intact hereditary units from damaged hereditary units. Thus, yeast cells ceased their cell division when they contained a chromosome without a telomere (Sandell and Zakian, 1993).

Telomeres fulfil another important task in association with the replication of eukaryotic cell DNA. In contrast to the circular genomes of prokaryotes, the linear chromosomes of eukaryotes cannot be completely replicated by the DNA polymerase complex. RNA primers are required to initiate DNA replication. After elimination of the RNA primers, extension of the Okazaki fragments and subsequent ligation, the newly synthesized DNA strand lacks the 5′ end since the RNA primer cannot be replaced by DNA at that point. Without special protective mechanisms, the chromosomes would therefore shrink with each cell division (“end-replication problem”; Harley et al., 1990). The non-coding telomere sequences presumably constitute a buffer-zone for preventing the loss of genes (Sandell and Zakian, 1993).

In addition to this, telomeres also play an import role in regulating cell ageing (Olovnikov, 1973). Human somatic cells exhibit a limited capacity for replication in culture; after a certain period of time, they become senescent. In this state, the cells no longer divide even after having been stimulated with growth factors; however, they do not die and remain metabolically active (Goldstein, 1990). Various observations support the hypothesis that a cell determines how many more times it can divide on the basis of the length of its telomeres (Allsopp et al., 1992).

In summary, the telomeres consequently possess key functions in the ageing of cells, and in stabilizing the genetic material and preventing cancer.

The Enzyme Telomerase Synthesizes the Telomeres

As described above, organisms which possess linear chromosomes can only replicate their genome incompletely in the absence of a special protective mechanism. Most eukaryotes use a special enzyme, i.e. telomerase, for regenerating the telomere sequences. Telomerase is expressed constitutively in the single-cell organisms which have so far been investigated. On the other hand, telomerase activity has only been measured in humans in germ cells and tumour cells, whereas neighbouring somatic tissue did not contain any telomerase (Kim et al., 1994).

Telomerase can also be designated functionally as terminal telomere transferase, which is located in the cell nucleus as a multiprotein complex. While the RNA moiety of human telomerase has been known for a relatively long, period of time (Feng et al., 1995), the catalytic subunit of this enzyme group was recently identified in a variety of organisms (Lingner et al., 1997; cf. our application PCT EP/98/03468 which is likewise pending). These catalytic subunits of telomerase are strikingly homologous both among themselves and in relation to all previously known reverse transcriptases.

WO 98/14592 also describes nucleic acid and amino acid sequences of the catalytic telomerase subunit.

Activation of Telomerase in Human Tumours

It was originally only possible to demonstrate telomerase activity in humans in germ line cells and not in normal somatic cells (Hastie el al., 1990; Kim et al., 1994). Following the development of a more sensitive detection method (Kim et al., 1994), a low telomerase activity was also detected in hematopoietic cells (Broccoli el al., 1995; Counter et al., 1995; Hiyama et al., 1995). It is true, however, that these cells nevertheless exhibited a reduction in the telomeres (Vaziri et al., 1994; Counter et al., 1995). It has still not been resolved whether the quantity of enzyme in these cells is not sufficient for compensating the telomere loss or whether the telomerase activity which is measured stems from a subpopulation, e.g. incompletely differentiated CD34⁺38⁺ precursor cells (Hiyama et al., 1995). In order to resolve this, it would be necessary to detect telomerase activity in a single cell.

Interestingly, however, significant telomerase activity was detected in a large number of the tumour tissues which had thus far been tested (1734/2031, 85%; Shay, 1997), whereas no activity was found in normal somatic tissue (1/196, <1%, Shay, 1997). In addition various investigations have shown that the telomeres still shrank in senescent cells which were transformed with viral oncoproteins and it was only possible to detect telomerase in the subpopulation which survived the growth crisis (Counter et al., 1992). The telomeres were also stable in these immortalized cells. (Counter et al., 1992). Similar findings from investigations in mice (Blasco et al., 1996) support the assumption that reactivation of the telomerase is a late event in tumorigenesis.

Based on these results, a “telomerase hypothesis” was developed which links the loss of telomere sequences and cell ageing with telomerase activity and the development of cancer. In long-lived species such as humans, the shrinking of the telomeres can be regarded as being a mechanism for suppressing tumours. Differentiated cells which do not contain any telomerase cease their cell division at a particular telomere length. If such a cell mutates, it can only form a tumour it the cell can extend its telomeres. Otherwise, the cell would continue to lose telomere sequences until its chromosomes became unstable and it was finally destroyed. Telomerase reactivation is presumably the main mechanism used by tumour cells to stabilize their telomeres.

It follows from these observations and considerations that it should be possible to treat tumours by inhibiting the telomerase. Conventional cancer therapies using cytostatic agents or short-wave radiation damage all the dividing cells in the body in addition to the tumour cells. However, since only germ line cells, apart from tumour cells, contain significant telomerase activity, telomerase inhibitors would attack the tumour cells more specifically and consequently elicit fewer undesirable side effects. Telomerase activity has been detected in all the tumour tissues which have so far been tested, which means that these therapeutic agents could be employed against all types of cancer. The effect of telomerase inhibitors would then set in when the telomeres of the cells had shortened to such an extent that the genome became unstable. Since tumour cells usually possess telomeres which are shorter than those of normal somatic cells, cancer cells would be the first to be eliminated by the telomerase inhibitors. By contrast, cells possessing long telomeres, such as the germ cells, would only be damaged at a much later date. Telomerase inhibitors consequently represent a potential way forward in the treatment of cancer.

It becomes possible to obtain unambiguous answers to the question of the nature and points of attack of physiological telomerase inhibitors once the manner in which expression of the telomerase gene is regulated has also been identified.

Regulation of Gene Expression in Eukaryotes

There are a large number of points in eukaryotic gene expression, i.e. the cellular flow of information from the DNA to the protein by way of the RNA, at which regulatory mechanisms can exert an effect. Examples of individual control steps are gene amplification the recombination of gene loci, chromatin structure, DNA methylation, transcription, post-transcriptional modifications of mRNA, mRNA transport, translation and post-translational modifications of proteins. Studies which have been carried out to date indicate that control at the level of transcription initiation is of the greatest importance (Latchman, 1991).

A region which is responsible for regulating transcription, and which is designated the promoter region, is located directly upstream of the transcription start of a gene which is transcribed by RNA polymerase II. Comparison of the nucleotide sequences of promoter regions from a large number of known genes shows that particular sequence motifs occur regularly in this region. These elements include, inter alia, the TATA box, the CCAAT box and the GC box, which elements are recognized by specific proteins. The TATA box, which is located about 30 nucleotides upstream of the transcription start, is, for example, recognized by the TFIID subunit TBP (“TATA box-binding protein”), whereas particular GC-rich sequence segments are specifically bound by the transcription factor Sp1 (“specificity protein 1”).

The promoter can be functionally subdivided into a regulatory segment and a constitutive segment (Latchman, 1991). The constitutive control region comprises the so-called core promoter switch enables transcription to be initiated correctly. This promoter contains the sequence elements which are described as UPE's (upstream promoter elements) which are necessary for efficient transcription. The regulatory control segments, which can be interlaced with the UPE's, possess sequence elements which can be involved in the signal-dependent regulation of transcription by hormones, growth factors, etc. They impart tissue-specific or cell-specific promoter properties.

DNA segments which are able to exert an influence on gene expression over relatively large distances are a characteristic feature of eukaryotic genes. These elements can be located upstream or downstream of a transcription unit, or within the unit, and can perform their function independently of their orientation. These sequence segments may reinforce (enhancers) or attenuate (silencers) promoter activity. In a similar way to the promoter regions, enhancers and silencers also accommodate several binding sites for transcription factors.

The invention relates to the DNA sequences from the 5′-flanking region of the gene for the catalytically active human telomerase subunit and intron sequences for this gene.

The invention particularly relates to the 5′-flanking regulatory DNA sequence which contains the promoter DNA sequence for the gene for the human catalytic telomerase subunit, as depicted in FIG. 10 (SEQ ID NO 3).

The invention furthermore relates to part regions of the 5′-flanking regulatory DNA sequence, as depicted in FIG. 4 (SEQ ID NO 1), which has a regulatory effect.

Intron sequences for the gene for the human catalytic telomerase subunit, in particular those sequences which have a regulatory effect, are also part of the subject-matter of the present invention. The intron sequences according to the invention are described in detail in the context of Example 5 (cf. SEQ ID NO 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20).

The invention furthermore relates to a recombinant construct which comprises the DNA sequences according to the invention, in particular the 5′-flanking DNA sequence of the gene for the human catalytic telomerase subunit, or part regions thereof.

Preference is given to recombinant constructs which, in addition to the DNA sequences according to the invention, in particular the 5′-flanking DNA sequence of the gene for the human catalytic telomerase subunit, or part regions thereof, also contain one or more additional DNA sequences which encode polypeptides or proteins.

According to a particularly preferred embodiment, these additional DNA sequences encode antineoplastic proteins.

Particular preference is given to those antineoplastic proteins which inhibit angiogenesis directly or indirectly. Examples of these proteins are:

Plasminogen activator inhibitor (PAI-1), PAI-2, PAI-3, angiostatin, endostatin, platelet factor 4, TIMP-1, TIMP-2, TIMP-3 and leukaemia inhibitory factor (LIF).

Antineoplastic proteins which have a direct or indirect cytostatic effect on tumours are likewise particularly preferred. These proteins include, in particular: perforin, granzyme, IL-2, IL-4,- IL-12, interferons, such as IFN-α, IFN-β and IFN-γ, TNF, TNF-α, TNF-β, oncostatin M; tumour suppressor genes, such as p53, retinoblastoma.

Particular preference is furthermore given to antineoplastic proteins which, where appropriate in addition to their antineoplastic effect, stimulate inflammations and thereby contribute to the elimination of tumour cells. Examples of these proteins are:

RANTES, monocyte chemotactic and activating factor (MCAF), IL-8, macrophage inflammatory protein (MIP-1α,-β), neutrophil activating protein-2 (NAP-2), IL-3, IL-5, human leukaemia inhibitory factor (LIF), IL-7, IL-I 1, IL-13, GM-CSF, G-CSF and M-CSF.

Particular preference is furthermore given to antineoplastic proteins which, due to their action as enzymes, are able to convert precursors of an antineoplastic active compound into an antineoplastic active compound Examples of these enzymes are:

herpes simplex virus thymidine kinase, varicella zoster virus thymidine kinase, bacterial nitroreductase, bacterial β-glucuronidase, plant β-glucuronidase from Secale cereale, human glucuronidase, human carboxy peptidase, bacterial carboxypeptidase, bacterial β-lactamnase, bacterial cytosine deaminidase, human catalase and/or phosphatase, human alkaline phosphatase, type 5 acid phosphatase, human lysooxidase, human acid D-aminooxidase, human glutathione peroxidase, human eosinophil peroxidase and human thyroid peroxidase.

The abovementioned recombinant constructs can also contain DNA sequences which encode factor VIII or factor IX, or part fragments thereof. These DNA sequences also include other blood clotting factors.

The abovementioned recombinant constructs can also contain DNA sequences which encode a reporter protein. Examples of these reporter proteins are:

Chloramphenicol acetyl transferase (CAT), glow-worm luciferase (LUC), β-galactosidase (β-Gal), secreted alkaline phosphatase (SEAP), human growth hormone (hGH), β-glucuronidase (GUS), green-fluorescing protein (GFP), and all the variants derived therefrom, aquarin and obelin.

Recombinant constructs according to the invention can also contain DNA which encodes the human catalytic telomerase subunit and its variants and fragments in the antisense orientation. Where appropriate, these constructs can also contain other protein subunits of the human telomerase and the telomerase RNA component in the antisense orientation.

The recombinant constructs can, in addition to the DNA which encodes the human catalytic telomerase subunit, and its variants and fragments, also contain other protein subunits of the human telomerase and the telomerase RNA component.

The invention furthermore relates to a vector which contains the abovementioned DNA sequences according to the invention, in particular the 5′-flanking DNA sequences and also one or more of the other DNA sequences mentioned above.

The preferred vector for these constructs is a virus, for example a retrovirus, an adenovirus, an adeno-associated virus, a herpes simplex virus, a vaccina virus, a lentiviral virus, a Sindbis virus and a Semliki forest virus.

Preference is also given to using plasmids as vectors.

The invention furthermore relates to pharmaceutical preparations which comprise recombinant constructs or vectors according to the invention; for example a preparation in a colloidal dispersion system.

Examples of suitable colloidal dispersion systems are liposomes or polylysine ligands.

The preparations of the constructs or vectors according to the invention in colloidal dispersion systems can be supplemented with a ligand which binds to the membrane structures of tumour cells. Such a ligand can, for example, be attached to the construct or the vector or else be a component of the liposome structure.

Suitable ligands are, in particular, polyclonal or monoclonal antibodies, or antibody fragments thereof, which bind, by their variable domains, to the membrane structures of tumour cells, or substances carrying mannose terminally, cytokines or growth factors, or fragments or part sequences thereof, which bind to receptors on tumour cells.

Examples of corresponding membrane structures are receptors for a cytokine or a growth factor, such as IL-1, EGF, PDGF, VEGF, TGF β, insulin or insulin-like growth factor (ILGF), or adhesion molecules, such as SLeX, LFA-1, MAC-1, LECAM-1 or VLA-4, or the mannose-6-phosphate receptor.

The present invention includes pharmaceutical preparations which, in addition to the vector constructs according to the invention, can also comprise non-toxic, inert, pharmaceutically suitable excipients. It is possible to conceive of administering (e.g. intravenously, intraarterially, intramuscularly, subcutaneously, intradermally, anally, vaginally, nasally, transdermally, intraperitoneally, as an aerosol or orally) these preparations at the site of a tumour or administering them systematically.

The vector constructs according to the invention can be employed in gene therapy.

The invention furthermore relates to a recombinant host cell, in particular a recombinant eukaryotic host cell, which harbours the above-described constructs or vectors.

The invention furthermore relates to a process for identifying substances which affect the promoter activity, silencer activity or enhancer activity of the catalytic telomerase subunit, with this process comprising the following steps:

-   -   A. adding a candidate substance to a host cell which harbours         the regulatory DNA sequence according to the invention, in         particular the 5′-flanking regulatory DNA sequence for the gene         for the human catalytic telomerase subunit, or a part region         thereof which has a regulatory effect, which sequence or part         region is functionally linked to a reporter gene, and     -   B. measuring the effect of the substance on expression of the         reporter gene.

The process can be employed for identifying substances which increase the promoter activity, silencer activity or enhancer activity of the catalytic telomerase subunit.

The process can furthermore be employed for identifying substances which inhibit the promoter activity, silencer activity or enhancer activator of the catalytic telomerase subunit.

The invention furthermore relates to a process for identifying factors which bind specifically to fragments of the DNA fragments according to the invention, in particular the 5′-flanking regulation DNA sequence of the catalytic telomerase subunit. This method comprises screening an expression cDNA library using the above-described DNA sequence, or subfragments of widely differing length, as the probe.

The above-described constructs or vectors can also be used for preparing transgenic animals.

The invention furthermore relates to a process for detecting telomerase-associated conditions in a patient, which process comprises the following steps:

-   -   A. incubating a construct or vector, which contains the DNA         sequence according to the invention, in particular the         5′-flanking regulatory DNA sequence for the gene for the human         catalytic telomerase subunit, or a part region thereof having a         regulatory effect, and a reporter gene, with body fluids or cell         samples,     -   B. detecting the activity of the reporter gene in order to         obtain a diagnostic value; and     -   C. comparing the diagnostic value with standard values for the         reporter gene construct in standardized normal cells or body         fluids of the same type as the test sample;

The detection of diagnostic values which are higher or lower than the standard comparative values indicates a telomerase-associated condition, which in turn indicates a pathogenic condition.

EXPLANATION OF THE FIGURES

FIG. 1: Southern blot analysis using genomic DNA from various species

-   -   A: Photograph of an ethidium bromide-stained 0.70% agarose gel         containing approximately 4 μg of Eco RI-cut (genomic DNA. Track         1 contains Hind III-cut λ DNA as size markers (23.5, 9.4, 6.7,         4.4, 2.3, 2.0 and 0.6 kb). Tracks 2 to 10 contain human, rhesus         monkey. Sprague Dawley rat, BALB/c mouse, dog, bovine, rabbit,         chicken and yeast (Saccharomyces cerevisiae) genomic DNA.     -   B: Autoradiogram, corresponding to FIG. 1 A, of a Southern blot         analysis in which radioactively labelled hTC-cDNA probe of about         720 bp in length is used for the hybridization.

FIG. 2: Restriction analysis of the recombinant λ DNA of the phage clone P12, which hybridizes with a probe from the 5′ region of the hTC cDNA.

The figure shows a photograph of an ethidium bromide-stained 0.4% agarose gel. Tracks 1 and 2 contain Eco RI/Hind III-cut λ DNA and a 1 kb ladder from Gibco as size markers. Tracks 3-7 each contain 250 ng of the DNA from the recombinant phage which has been cut with Bam HI (track 3), Eco RI (track 4), Sal I (track 5), Xho I (track 6) and Sac I (track 7). The arrows mark the two λ arms of the vector EMBL3 Sp6/T7.

FIG. 3: Restriction analysis and Southern blot analysis of the recombinant λ DNA of the phage clone which hybridizes with a probe from the 5′ region of the hTC cDNA.

-   -   A: The figure shows a photograph of an ethidium bromide-stained         0.8% agarose gel. Tracks 1 and 15 contain a 1 kb ladder from         Gibco as size markers. Tracks 2 to 14 each contain 250 ng of cut         λ DNA from the recombinant phage clone. The following enzymes         were employed: track 2: Sac I, track 3: Xho I, track 4: Xho I,         Xba I, track 5: Sac I, Xho I, track 6: Sal I, Xho I, Xba I,         track 7: Sac I, Xho I, Xba I, track 8: Sac I, Sal I, Xba I,         track 9: Sac I, Sal I, BamH I, track 10: Sac I, Sal I, Xho I,         track 11: Not I, track 12: Sma I, track 13: empty. track 14: not         digested.     -   B: Autoradiogram, corresponding to FIG. 3 A, of a Southern blot         analysis. A 5′-hTC cDNA fragment of about 420 bp in length was         used as the probe for the hybridization.

FIG. 4: Partial DNA sequence of the 5′-flanking region and of the promoter of the gene for the human catalytic telomerase subunit. The ATG start codon in the sequence is printed in bold. The depicted sequence corresponds to SEQ ID. NO 1.

FIG. 5: Use of primer extension analysis to identify the transcription start.

The figure shows an autoradiogram of a denaturing polyacrylamide gel which was selected for depicting a primer extension analysis. An oligonucleotide having the sequence 5′GTTAAGTTGTAGCTTACACTGGTTCTC 3′ was used as the primer. The primer extension reaction was loaded in track 1. Tracks G, A, T and C constitute the sequence reactions using the same primer and the corresponding dideoxynucleotides. The thick arrow marks the main transcription start while the thin arrows point to three subsidiary transcription start points.

FIG. 6: cDNA sequence of the human catalytic telomerase subunit (hTC; cf. our pending application PCT/EP/98/03468). The depicted sequence corresponds to SEQ ID NO 2.

FIG. 7: Structural organization and restriction map of the human hTC gene and its 5′-flanking and 3′-flanking regions.

Exons are shown as consecutively numbered rectangles which are filled-in in black and introns are shown as regions which are not filled in. Untranslated sequence segments in the exons are hatched. Translation starts in exon 1 and ends in exon 16. Restriction enzyme cleavage sites are marked as follows: S, SacI; X, XhoI. The relative arrangement of the five phage clones (P2, P3, P5, P12, P17), and of the product from the genome walking, are shown by thin lines. As the dots indicate, the sequence of intron 16 has only been partly deciphered.

FIG. 8: HTL splice variants.

-   -   A: Diagrammatic structure of the hTC mRNA splice variants. The         complete hTC mRNA is depicted as a rectangle with a grey         background in the upper region of the figure. The 16 exons are         depicted in accordance with their size. The translation start         (ATG) and the stop codon, and also the telomerase-specific T         motif, and the seven RT motifs, are all shown. The hTC variants         are subdivided into deletion and insertion variants. The missing         exon sequences are marked in the deletions. The insertions are         shown by additional white rectangles. The sizes and origins of         the inserted sequences are given. Newly formed stop codons are         marked. The size of the insertion in variant INS2 is unknown.     -   B: Exon-intron transitions in the hTC splice variants. Unspliced         5′-flanking and 3′-flanking sequences are shown as white         rectangles. The origins of the exon and intron sequences are         given. Intron and exon sequences are shown in small letters and         large letters, respectively. The donor and acceptor sequences in         the splice sites are underlaid as grey rectangles, and their         exon and intron origins are also given.

FIG. 9: Identification of the transcription start by means of RT-PCR analysis. The RT-PCR was carried out using a cDNA library prepared from HL 60 cells and genomic DNA as the positive control. A common 3′ primer hybridizes to a region of the exon I sequence. The positions of the different 5′ primers in the coding region or the 5′-flanking region are given. In the negative control, no template DNA was added to the PCR reaction. M: DNA size marker.

FIG. 10: Nucleotide sequence and structural features of the hTC promoter.

The figure depicts 11273 bp of the 5′-flanking hTC gene sequence, beginning with the translation start codon ATG (+1). The putative region of the translation start is underlined. Possible regulatory sequence segments within the 4000 bp upstream of the translation start are ringed. The depicted sequence corresponds to SEQ ID NO 3.

FIG. 11: Activity of the hTC promoter in HEK-293 cells.

The first 5000 bp of the 5′-flanking hTC gene region are shown diagrammatically in the upper part of the figure. The ATG start codon is picked out. CpG-rich islands are marked by grey rectangles. The sizes of the hTC promoter-luciferase construct are shown on the left-hand side of the figure. The promoterless. pGL2 basic construct and the SV40 promoter construct pGL2-Pro were used as controls in each transfection. The relative luciferase activities of the different promoter constructs in HEK cells are shown as continuous bars on the right-hand side of the figure. The standard deviation is indicated. The numerical values represent the average of two independent experiments which were carried out in duplicate.

Tab. 1: Exon-intron transitions in the hTC gene

The table lists the nucleotide sequences at the 3′ and 5′ splice transitions of the hTC gene. The consensus sequences for donor and acceptor sequences (AG and GT) are underlaid with grey rectangles. The table shows the intron sequences (small letters) and exon sequences (large letters) which flank the splice acceptor and donor sites. The sizes of the exons and introns are given in bp.

Tab. 2: Potential binding sites for DNA-binding factors in the nucleotide sequence of intron 2

The search for possible DNA-binding factors (e.g. transcription factors) was carried out using the “find pattern” algorithm from the Genetics Computer Group (Madison, USA) GCG sequence analysis program package. The table lists the abbreviations of the DNA-binding factors which were identified and their location in intron 2. TAB. 1

TAB 2 Factors Location in intron 2 C/EBP 2925 CRE.2 2749 Sp1 2378, 4094, 4526, 4787, 4835, 4995 AP-2 CS3 5099 AP-2 CS4 2213, 3699, 4667, 5878, 5938, 6059, 6180, 6496 AP-2 CS5 5350, 5798, 5880, 5940, 6061, 6182, 6375, 6498 PEA3 934, 2505 P53 2125 GR uteroglobin 848, 1487, 2956 PR uteroglobin 3331 Zeste-white 1577, 1619, 1703, 1745, 1787, 1829, 1871, 1913, 1955, 1997, 2039, 2081, 3518, 3709, 4765, 5014, 5055 GRE 846 MyoD-MCK 447, 509, 558, 1370, 1595, 1900, 2028, 2099, 4557 right site/rev MyoD-MCK 108, 118, 453, 1566, 1608, 1692, 1734, 1818, 1902, left site 1986, 2372, 2460, 2720, 3491, 5030 Ets-1 CS 6408 AP1 3784, 4406 CREB 2801 GATA-1 839, 1390, 3154 c-Myc 108, 118, 453, 1566, 1608, 1692, 1734, 1818, 1902, 1986, 2372, 2460, 2720, 3491, 5030 CACCC site 991 CCAAT site 1224 CCAC box 992 CAAT site 463, 2395 Rb site 992, 4663 TATA 3650 CDEI 106, 1564, 1606, 1690, 1732, 1816, 1900, 1984

EXAMPLES

The human gene for the catalytic telomerase subunit (ghTC), and the regions of this gene located 5′ and 3′, were cloned, while the start point for transcription was determined, potential binding sites for DNA-binding proteins were identified and active promoter fragments were highlighted. The sequence of the hTC cDNA (FIG. 6) has already been reported in our application PCT/EP/98/03468, which is also pending. Unless otherwise mentioned, all the data refer to the position of the cDNA in this sequence.

Example 1

A genomic Southern blot analysis was used to determine whether ghTC constitutes a single gene in the human genome or whether there exist several loci for the hTC gene and possibly also ghTC pseudogenes.

In order to do this, a commercially available zoo blot from Clontech was subjected to Southern blot analysis. This blot contains 4 μg of Eco RI-cut genomic DNA from nine different species (human, monkey, rat, mouse, dog, bovine, rabbit, chicken and yeast). With the exception of yeast, chicken and human, the DNA was isolated from kidney tissue. The human genomic DNA was isolated from placenta and the chicken genomic DNA was purified from liver tissue. An hTC cDNA fragment of about 720 bp in length, which was isolated from hTC cDNA, variant Del2 (position 1685 to 2349 plus 2531 to 2590 in FIG. 6 [deletion 2; cf. Example 5 in FIG. 8]), was used as the radioactively labelled probe in the autoradiogram in FIG. 1. The experimental conditions for the blot hybridization and washing steps were taken from Ausubel et al. (1987).

In the case of the human DNA, the probe recognizes two specific DNA fragments. The smaller Eco RI fragment, of from about 1.5 to 1.8 kb in length, probably originates from two Eco RI cleavage sites in an intron in the ghTC DNA. On the basis of this result, it is to be assumed that only one single ghTC gene is present in the human genome.

Example 2

In order to isolate the 5′ flanking hTC gene sequence, approx. 1.5×10⁶ phages from a human genomic placenta gene library (EMBL 3 SP6/T7 from Clontech, order number HL1067j) were hybridized on nitrocellulose filters (0.45 μm; from Schleicher and Schuell), in accordance with the manufacturer's instructions, with a radioactively labelled 5′-hTC cDNA fragment of about 500 bp in length (position 839 to 1345 in FIG. 6). The nitrocellulose filters were firstly incubated, at 42° C. for two hours, in 2×SSC (0.3 M NaCl; 0.5 M Tris-HCl, pH 8.0) and then in a prehybridization solution (50% formamide; 5×SSPE, pH 7.4; 5×Denhard's solution; 0.25% SDS; 100 μg of herring sperm DNA/ml). For the overnight hybridization, the prehybridization solution was supplemented with 1.5×10⁶ cpm of denatured, radioactively labelled probe/ml of solution. Nonspecifically bound radioactive DNA was removed under stringent conditions, i.e. by means of three five-minute steps of washing with 2×SSC; 0.1% SDS at from 55 to 65° C. The filters were evaluated by autoradiography.

The phage clones which were identified in this primary investigation were purified (Ausubel et al. (1987)). In subsequent analyses, one phage clone, i.e. P12 turned out to be potentially positive. A λ DNA preparation carried out on this phage (Ausubel et al. (1987)), and the subsequent restriction digestion with enzymes which release the genomic insert in fragments, showed that this phage clone contains an insert of approx. 15 kb in the vector (FIG. 2).

In order to isolate the complete hTC gene sequence, in each case from 1 to 1.5×10⁶ phages were screened, in independent experiments, with in each case different radioactively labelled probes, as described above.

The phage clones which were identified in these primary investigations, and which were positive for the corresponding probes, were purified. The phage clone P17 was found to contain an hTC cDNA fragment of about 250 bp in length (position 1787 to 2040 in FIG. 6). The phage clone P2 was identified as containing an hTC cDNA fragment of about 740 bp in length (position 1685 to 2349 plus 2531 to 2607 in FIG. 6 [deletion 2; cf. Example 5]). The phage clones P3 and P5 were found to contain a 3′ hTC cDNA fragment of 420 bp in length (position 3047 to 3470 in FIG. 6). After the λ DNA had been prepared from these phages, and subsequently subjected to restriction digestion with enzymes which release the genomic insert in fragments, the inserts were subcloned into plasmids (Example 4).

Example 3

In order to investigate whether the 5′ end of the hTC cDNA was also present in the insert in the recombinant phage clone P12, the λ DNA from this clone was hybridized, in a Southern blot analysis, with a radiactively labelled hTC cDNA fragment of about 440 bp in length (position 1 to 440 in FIG. 6) from the extreme 5′ region (FIG. 3).

Since the isolated λ DNA from the positive clone also hybridizes with the extreme 5′ end of the hTC cDNA, this phage probably also contains the 5′ sequence region flanking the ATG start codon.

Example 4

In order to subclone the entire 15 kb insert in the positive phage clone P12 in the form of subfragments, and subsequently to sequence these fragments, restriction endonucleases which, on the one hand, release the entire insert from EMBL3 Sp6/T7 (cf. Example 2) and, in addition, cut within the insert, were selected for digesting the DNA.

In all, two Xho I subfragments, of about 8.3 and about 6.5 kb in length, respectively, and three Sac I subfragments, of about 8.5, about 3.5 and about 3 kb in length, respectively, were subcloned into the pBluescript KS(+) vector (from Stratagene). The 5123 bp 5′-flanking nucleotide sequence of the ghTC gene region, starting from the ATG start codon, was determined by analysing the sequences of these fragments (FIG. 4; corresponding to SEQ ID NO 1). FIG. 4 depicts the first 5123 bp (starting from the ATG start codon). FIG. 10 depicts the entire cloned 5′ sequence (corresponding to SEQ ID NO 3).

In order to subclone the entire insert, of approx. 14.6 kb in size, in phage clone P17 in the form of subfragments, restriction endonucleases which, on the one hand, release the entire insert from EMLB3 Sp6/T7 and, in addition, cut a few times within the insert, were selected for digesting the DNA. Three XhoI/BamHI fragments, of 7.1 kb, 4.2 kb and 1.5 kb in size, respectively, and one BamHI fragment, of 1.8 kb in size, were subcloned by means of using a combination digestion with the enzymes XhoI and BamHI. Combination restriction digestion with the enzymes XhoI and XbaI resulted in a XhoI/XbaI fragment of 6.5 kb in size, and two XhoI fragments, of 6.5 kb and 1.5 kb in size, respectively, being cloned.

Digestion with the restriction enzyme XhoI was used to subclone the insert, of approx. 17.9 kb in size, in phage clone P2 in the form of subfragments. In all, three XhoI subfragments, of 7.5 kb, 6.4 kb and 1.6 kb in length, respectively, were cloned. Four SacI fragments, of 4.8 kb, 3 kb, 2 kb and 1.8 kb in size, respectively, were additionally subcloned by digesting with the restriction enzyme SacI.

The insert, of approx. 13.5 kb in size, in phage clone P3 was subcloned by digesting with the restriction enzymes SacI and/or XhoI. Six SacI subfragments, of 3.2 kb, 2 kb, 0.9 kb, 0.8 kb, 0.65 kb and 0.5 kb in length, respectively, and two XhoI subfragments, of 6.5 kb and 4.3 kb in length, respectively, were obtained in this connection.

The insert, of approx. 13.2 kb in size, in phage clone P5 was subcloned by digesting with the restriction enzymes SacI and/or XhoI. In all, SacI fragments of 6.5 kb, 3.3 kb, 3.2 kb, 0.8 kb and 0.3 kb in size, and Xhol fragments of 7 kb and 3.2 kb in size, were subcloned.

In order to clone the hTC genomic sequence region located 3′ of phage clone P17 and 5′ of phage clone P2, 3 genomic walkings were carried out using the Clontech GenomeWalker™ kits (catalogue number K1803-1) and various combinations of primers. In a final volume of 50 μl, 10 pmol of dNTP mix were added to 1 μl of human GenomeWalker Library HDL (from Clontech), and a PCR reaction was carried out in 1×Klen Taq PCR reaction buffer and 1×Advantage Klen Taq polymerase mix (from Clontech). 10 pmol of an internal gene-specific primer, and 10 pmol of the adaptor primer AP1 (5′-GTAATACGACTCACTATAGGGC-3′; from Clontech) were added as primers. The PCR was carried out in 3 steps as a touchdown PCR. First of all, denaturation was carried out at 94° C. for 20 sec, and the primers were then annealed, and the DNA chain extended, at 72° C. for 4 min, over 7 cycles. There then followed 37 cycles in which the DNA was denaturated at 94° C. for 20 sec but the subsequent primer extension took place at 67° C. for 4 min. In conclusion, there followed a chain extension at 67° C. for 4 min. After this first PCR, the PCR product was diluted 1:50. One μl of this dilution- was used in a second nested PCR together with 10 pmol of dNTP mix in 1×Klen Taq PCR reaction buffer and 1×Advantage KIen Taq polymerase mix and also 10 pmol of a nested gene-specific primer and 10 pmol of the nested Marathon Adaptor primers AP2 (5′-ACTATAGGGCACGCGTGGT-3′; from Clontech). The PCR conditions corresponded to the parameters which were selected in the first PCR. As the sole exception, only 5 cycles rather than 7 cycles were selected in the first PCR step and only 24 cycles, instead of 37 cycles. were run in the second PCR step. The products of this nested genomic walking PCR were cloned into the TA Cloning Vector pCRII from InVitrogen.

In the first genomic walking, the gene-specific primer C3K2-GSP1 (5′-GACGTGGCTCTTGAAGGCCTTG-3′) and the nested gene-specific primer C3K2-GSP2 (5′-GCCTTCTGGACCACGGCATACC-3′) were used, together with the HDL library 4, and a PCR fragment of 1639 bp in length was obtained. In the second genomic walking, a PCR fragment of 685 bp in length was amplified from the HDL library 4 using the gene-specific primer C3F2 (5′-CGTAGTTGAGCACGCTGAACAGTG-3′) and the nested gene-specific primer C3F (5′-CCTTCACCCTCGAGGTGAGACGCT-3. The third genomic walking mixture, using the gene-specific primer DEL5-GSP1 (5′-GGTGGATGTGACGGGCGCGTACG-3′) and the nested gene-specific primer C5K-GSP1 (5′-GGTATGCCGTGGTCCAGAAGGC-3′), led to a 924 bp PCR fragments being cloned from the HDL library 1. In all, 2100 bp of the genomic hTC region located 3′ of phage clone P17 were identified using this genomic walking method (see FIG. 7).

The subcloned fragments, and the genomic walking products, were sequenced in single-stranded form. The Lasergene Biocomputing Software (DNASTAR Inc. Madison, Wis., USA) was used to identify overlapping regions and form contigs. In all, 2 large contigs were assembled from the sequences collected from phage clones P12, P17, P2, P3 and P5, and also the sequence data from the genomic walking. Contig 1 consists of sequence data from phage clones P12 and P17 and the sequence data from the genomic walking. Contig 2 was put together from the sequences from phage clones P2, P3 and P5. Overlapping phage clone regions are shown diagrammatically in FIG. 7. The sequence data from the 2 contigs are shown below. The ATG start codon in contig 1 is underlined. The TGA stop codon is underlined in contig 2. Contig 1: ACTTGAGCCC AAGAGTTCAA GGCTACGGTG AGCCATGATT GCAACACCAC ACGCCAGCCT TGGTGACAGA 70 ATGAGACCCT GTCTCAAAAA AAAAAAAAAA AATTGAAATA ATATAAAGCA TCTTCTCTGG CCACAGTGGA 140 ACAAAACCAG AAATCAACAA CAAGAGGAAT TTTGAAAACT ATACAAACAC ATGAAAATTA AACAATATAC 210 TTCTGAATGA CCAGTGAGTC AATGAAGAAA TTAAAAAGGA AATTGAAAAA TTTATTTAAG CAAATGATAA 280 CGGAAACATA ACCTCTCAAA ACCCACGGTA TACAGCAAAA GCAGTGCTAA GAAGGAAGTT TATAGCTATA 350 AGCAGCTACA TCAAAAAAGT AGAAAAGCCA GGCGCAGTGG CTCATGCCTG TAATCCCAGC ACTTTGGGAG 420 GCCAAGGCGG GCAGATCGCC TGAGGTCAGG AGTTCGAGAC CAGCCTGACC AACACAGAGA AACCTTCTCG 490 CTACTAAAAA TACAAAATTA GCTGGGCATG GTGGCACATG CCTGTAATCC CAGCTACTCG GGAGGCTGAG 560 GCAGGATAAC CGCTTGAACC CAGGAGGTGG AGGTTGCGGT GAGCCGGGAT TGCGCCATTG GACTCCAGCC 630 TGGGTAACAA GAGTGAAACC CTGTCTCAAG AAAAAAAAAA AAGTAGAAAA ACTTAAAAAT ACAACCTAAT 700 GATGCACCTT AAAGAACTAG AAAAGCAAGA GCAAACTAAA CCTAAAATTG GTAAAAGAAA AGAAATAATA 770 AAGATCAGAG CAGAAATAAA TGAAACTGAA AGATAACAAT ACAAAAGATC AACAAAATTA AAAGTTGGTT 840 TTTTGAAAAG ATAAACAAAA TTGACAAACC TTTGCCCAGA CTAAGAAAAA AGGAAAGAAG ACCTAAATAA 910 ATAAAGTCAG AGATGAAAAA AGAGACATTA CAACTGATAC CACAGAAATT CAAAGGATCA CTAGAGGCTA 980 CTATGAGCAA CTGTACACTA ATAAATTGAA AAACCTAGAA AAAATAGATA AATTCCTAGA TGCATACAAC 1050 CTACCAAGAT TGAACCATGA AGAAATCCAA AGCCCAAACA GACCAATAAC AATAATGGGA TTAAAGCCAT 1120 AATAAAAAGT CTCCTAGCAA AGAGAAGCCC AGGACCCAAT GGCTTCCCTG CTGGATTTTA CCAATCATTT 1190 AAAGAAGAAT GAATTCCAAT CCTACTCAAA CTATTCTGAA AAATAGAGGA AAGAATACTT CCAAACTCAT 1260 TCTACATGGC CAGTATTACC CTGATTCCAA AACCAGACAA AAACACATCA AAAACAAACA AACAAAAAAA 1330 CAGAAAGAAA GAAAACTACA GGCCAATATC CCTGATGAAT ACTGATACAA AAATCCTCAA CAAAACACTA 1400 GCAAACCAAA TTAAACAACA CCTTCGAAAG ATCATTCATT GTGATCAAGT GGGATTTATT CCAGGGATGG 1470 AAGGATGGTT CAACATATGC AAATCAATCA ATGTGATACA TCATCCCAAC AAAATGAAGT ACAAAAACTA 1540 TATGATTATT TCACTTTATG CAGAAAAAGC ATTTGATAAA ATTCTGCACC CTTCATGATA AAAACCCTCA 1610 AAAAACCAGG TATACAAGAA ACATACAGGC CAGGCACAGT GGCTCACACC TGCGATCCCA GCACTCTGGG 1680 AGGCCAAGGT GGGATGATTG CTTGGGCCCA GGAGTTTGAG ACTAGCCTGG GCAACAAAAT GAGACCTGGT 1750 CTACAAAAAA CTTTTTTAAA AAATTAGCCA GGCATGATGG CATATGCCTG TAGTCCCAGC TAGTCTGGAG 1820 GCTGAGGTGG GAGAATCACT TAAGCCTAGG AGGTCGAGGC TGCAGTGAGC CATGAACATG TCACTGTACT 1890 CCAGCCTAGA CAACAGAACA AGACCCCACT GAATAAGAAG AAGGAGAAGG AGAAGGGAGA AGGGAGGGAG 1960 AAGGGAGGAG GAGGAGAAGG AGGAGGTGGA GGAGAAGTGG AAGGGGAAGG GGAAGGGAAA GAGGAAGAAG 2030 AAGAAACATA TTTCAACATA ATAAAAGCCC TATATGACAG ACCGAGGTAG TATTATGAGG AAAAACTGAA 2100 AGCCTTTCCT CTAAGATCTG GAAAATGACA AGGGCCCACT TTCACCACTG TGATTCAACA TAGTACTAGA 2170 AGTCCTAGCT AGAGCAATCA GATAAGAGAA AGAAATAAAA GGCATCCAAA CTGGAAAGGA AGAAGTCAAA 2240 TTATCCTGTT TGCAGATGAT ATGATCTTAT ATCTGGAAAA GACTTAAGAC ACCACTAAAA AACTATTAGA 2310 GCTGAAATTT GGTACAGCAG GATACAAAAT CAATGTACAA AAATCAGTAG TATTTCTATA TTCCAACAGC 2380 AAACAATCTG AAAAAGAAAC CAAAAAAGCA GCTACAAATA AAATTAAACA GCTAGGAATT AACCAAAGAA 2450 GTGAAAGATC TCTACAATGA AAACTATAAA ATGTTGATAA AAGAAATTGA AGAGGGCACA AAAAAAGAAA 2520 AGATATTCCA TGTTCATAGA TTGGAAGAAT AAATACTGTT AAAATGTCCA TACTACCCAA AGCAATTTAC 2590 AAATTCAATG CAATCCCTAT TAAAATACTA ATGACGTTCT TCACAGAAAT AGAAGAAACA ATTCTAAGAT 2660 TTGTACAGAA CCACAAAAGA CCCAGAATAG CCAAAGCTAT CCTGACCAAA AAGAACAAAA CTGGAAGCAT 2730 CACATTACCT GACTTCAAAT TATACTACAA AGCTATAGTA ACCCAAACTA CATGGTACTG GCATAAAAAC 2800 AGATGAGACA TGGACCAGAG GAACAGAATA GAGAATCCAG AAACAAATCC ATGCATCTAC AGTGAACTCA 2870 TTTTTGACAA AGGTGCCAAG AACATACTTT GGGGAAAAGA TAATCTCTTC AATAAATGGT GCTGGAGGAA 2940 CTGGATATCC ATATGCAAAA TAACAATACT AGAACTCTGT CTCTCACCAT ATACAAAAGC AAATCAAAAT 3010 GGATGAAAGG CTTAAATCTA AAACCTCAAA CTTTGCAACT ACTAAAAGAA AACACCGGAG AAACTCTCCA 3080 GGACATTGGA GTGGGCAAAG ACTTCTTGAG TAATTCCCTG CAGGCACAGG CAACCAAAGC AAAAACAGAC 3150 AAATGGGATC ATATCAAGTT AAAAAGCTTC TGCCCAGCAA AGGAAACAAT CAACAAAGAG AAGAGACAAC 3220 CCACAGAATG GGAGAATATA TTTGCAAACT ATTCATCTAA CAAGGAATTA ATAACCAGTA TATATAAGGA 3290 GCTCAAACTA CTCTATAAGA AAAACACCTA ATAAGCTGAT TTTCAAAAAT AAGCAAAAGA TCTGGGTAGA 3360 CATTTCTCAA AATAAGTCAT ACAAATGGCA AACAGGCATC TGAAAATGTG CTCAACACCA CTGATCATCA 3430 GAGAAATGCA AATCAAAACT ACTATGAGAG ATCATCTCAT CCCAGTTAAA ATGGCTTTTA TTCAAAAGAC 3500 AGGCAATAAC AAATGCCAGT GAGGATGTGG ATAAAAGGAA ACCCTTGGAC ACTGTTGGTG GGAATGGAAA 3570 TTGCTACCAC TATGGAGAAC AGTTTGAAAG TTCCTCAAAA AACTAAAAAT AAAGCTACCA TACAGCAATC 3640 CCATTGCTAG GTATATACTC CAAAAAAGGG AATCAGTGTA TCAACAAGCT ATCTCCACTC CCACATTTAC 3710 TGCAGCACTG TTCATAGCAG CCAAGGTTTG GAAGCAACCT CAGTGTCCAT CAACAGACGA ATGGAAAAAG 3780 AAAATGTGGT GCACATACAC AATGGAGTAC TACGCAGCCA TAAAAAAGAA TGAGATCCTG TCAGTTGCAA 3850 CAGCATGGGG GGCACTGGTC AGTATGTTAA GTGAAATAAG CCAGGCACAG AAAGACAAAC TTTTCATGTT 3920 CTCCCTTACT TGTGGGAGCA AAAATTAAAA CAATTGACAT AGAAATAGAG GAGAATGGTG GTTCTAGAGG 3990 GGTGGGGGAC AGGGTGACTA GAGTCAACAA TAATTTATTG TATGTTTTAA AATAACTAAA AGAGTATAAT 4060 TGGGGTGTTT GTAACACAAA GAAAGGATAA ATGCTTGAAG GTGACAGATA CCCCATTTAC CCTGATGTGA 4130 TTATTACACA TTGTATGCCT GTATCAAAAT ATCTCATGTA TGCTATAGAT ATAAACCCTA CTATATTAAA 4200 AATTAAAATT TTAATGGCCA GGCACGGTGG CTCATGTCCG TAATCCCAGC ACTTTGGGAG GCCGAGGCGG 4270 GTGGATCACC TGAGGTCAGG AGTTTGAAAC CAGTCTGGCC ACCATGATGA AACCCTGTCT CTACTAAAGA 4340 TACAAAAATT AGCCAGGCGT GGTGGCACAT ACCTGTAGTC CCAACTACTC AGGAGCCTGA GACAGGAGAA 4410 TTGCTTGAAC CTGGGAGGCG GAGGTTGCAG TGAGCCGAGA TCATGCCACT GCACTGCAGC CTGGGTGACA 4480 GAGGAAGACT CCATCTCAAA ACAAAAACAA AAAAAAGAAG ATTAAAATTG TAATTTTTAT GTACCGTATA 4550 AATATATACT CTACTATATT AGAAGTTAAA AATTAAAACA ATTATAAAAG GTAATTAACC ACTTAATCTA 4620 AAATAAGAAC AATGTATGTG GGGTTTCTAG CTTCTGAAGA AGTAAAAGCT ATGGCCACGA TGGCAGAAAT 4690 GTGAGGAGGG AACAGTGGAA GTTACTGTTG TTAGACGCTC ATACTCTCTG TAAGTGACTT AATTTCAACC 4760 AAAGACAGGC TGGGAGAAGT TAAAGAGGCA TTCTATAAGC CCTAAAACAA CTGCTAATAA TGGTGAAAGG 4830 TAATCTCTAT TAATTACCAA TAATTACAGA TATCTCTAAA ATCGACCTGC AGAATTGGCA CGTCTCATCA 4900 CACCGTCCTC TCATTCACGG TGCTTTTTTT CTTGTGTGCT TGGAGATTTT CGATTGTGTG TTCGTGTTTG 4970 GTTAAACTTA ATCTGTATGA ATCCTGAAAC GAAAAATGGT GGTGATTTCC TCCAGAAGAA TTAGAGTACC 5040 TGGCAGGAAG CAGGTGGCTC TGTGGACCTG AGCCACTTCA ATCTTCAAGG GTCTCTGGCC AAGACCCAGG 5110 TGCAAGGCAG AGGCCTGATG ACCCGAGGAC AGGAAAGCTC GGATGGGAAG GGGCGATGAG AAGCCTGCCT 5180 CGTTGGTGAG CAGCGCATGA AGTGCCCTTA TTTACGCTTT GCAAAGATTG CTCTGGATAC CATCTGGAAA 5250 AGGCGGCCAG CGGGAATGCA AGGAGTCAGA AGCCTCCTGC TCAAACCCAG GCCAGCAGCT ATGGCGCCCA 5320 CCCGGGCGTG TGCCAGAGGG AGAGGAGTCA AGGCACCTCG AAGTATGGCT TAAATCTTTT TTTCACCTGA 5390 AGCAGTGACC AAGGTGTATT CTGAGGGAAG CTTGAGTTAG GTGCCTTCTT TAAAACAGAA AGTCATGGAA 5460 GCACCCTTCT CAAGGGAAAA CCAGACGCCC GCTCTGCGGT CATTTACCTC TTTCCTCTCT CCCTCTCTTG 5530 CCCTCGCGGT TTCTGATCGG GACAGAGTGA CCCCCGTGGA GCTTCTCCGA GCCCGTGCTG AGGACCCTCT 5600 TGCAAAGGGC TCCACAGACC CCCGCCCTGG AGAGAGGAGT CTGAGCCTGG CTTAATAACA AACTGGGATC 5670 TGGCTGGGGG CGGACAGCGA CGGCGGGATT CAAAGACTTA ATTCCATGAG TAAATTCAAC CTTTCCACAT 5740 CCGAATGGAT TTGGATTTTA TCTTAATATT TTCTTAAATT TCATCAAATA ACATTCAGGA CTGCAGAAAT 5810 CCAAAGGCGT AAAACAGGAA CTGAGCTATG TTTGCCAAGG TCCAAGGACT TAATAACCAT GTTCAGAGGG 5880 ATTTTTCGCC CTAAGTACTT TTTATTGGTT TTCATAAGGT GGCTTAGGGT GCAAGGGAAA GTACACGAGG 5950 AGAGGCCTGG GCGGCAGGGC TATGAGCACG GCAGGGCCAC CGGGGAGAGA GTCCCCGGCC TGGGAGGCTG 6020 ACAGCAGGAC CACTGACCGT CCTCCCTGGG AGCTGCCACA TTGGGCAACG CGAAGGCGGC CACGCTGCGT 6090 GTGACTCAGG ACCCCATACC GGCTTCCTGG GCCCACCCAC ACTAACCCAG GAAGTCACGG AGCTCTGAAC 6160 CCGTGGAAAC GAACATGACC CTTGCCTGCC TGCTTCCCTG GGTGGGTCAA GGGTAATGAA GTGGTGTGCA 6230 GGAAATGGCC ATGTAAATTA CACGACTCTG CTGATGGGGA CCGTTCCTTC CATCATTATT CATCTTCACC 6300 CCCAAGGACT GAATGATTCC AGCAACTTCT TCGGGTGTGA CAAGCCATGA CAAAACTCAG TACAAACACC 6370 ACTCTTTTAC TAGGCCCACA GAGCACGGSC CACACCCCTG ATATATTAAG AGTCCAGGAG AGATGAGGCT 6440 GCTTTCAGCC ACCAGGCTGG GGTGACAACA GCGGCTGAAC AGTCTGTTCC TCTAGACTAG TAGACCCTGG 6510 CAGGCACTCC CCCAGATTCT AGGGCCTGGT TGCTGCTTCC CGAGGGCGCC ATCTGCCCTG GAGACTCAGC 6580 CTGGGGTGCC ACACTGAGGC CAGCCCTGTC TCCACACCCT CCGCCTCCAG GCCTCAGCTT CTCCAGCAGC 6650 TTCCTAAACC CTGGGTGGGC CGTGTTCCAG CGCTACTGTC TCACCTGTCC CACTGTGTCT TGTCTCAGCG 6720 ACGTAGCTCG CACGGTTCCT CCTCACATGG GGTGTCTGTC TCCTTCCCCA ACACTCACAT GCGTTGAAGG 6790 GAGGAGATTC TGCGCCTCCC AGACTGGCTC CTCTGAGCCT GAACCTGGCT CGTGGCCCCC GATGCAGGTT 6860 CCTGGCGTCC GGCTGCACGC TGACCTCCAT TTCCAGGCGC TCCCCGTCTC CTGTCATCTG CCGGGGCCTG 6930 CCGGTGTGTT CTTCTGTTTC TGTGCTCCTT TCCACCTCCA GCTGCGTGTG TCTCTGCCCG CTAGGGTCTC 7000 GGGGTTTTTA TAGGCATAGG ACGGGGGCGT GGTGGGCCAG GGCGCTCTTG GGAAATGCAA CATTTGGGTG 7070 TGAAAGTAGG AGTGCCTGTC CTCACCTAGG TCCACGGGCA CAGGCCTGGG GATGGAGCCC CCGCCAGGGA 7140 CCCGCCCTTC TCTGCCCAGC ACTTTCCTGC CCCCCTCCCT CTGGAACACA GAGTGGCAGT TTCCACAAGC 7210 ACTAAGCATC CTCTTCCCAA AAGACCCAGC ATTGGCACCC CTGGACATTT GCCCCACAGC CCTGGGAATT 7280 CACGTGACTA CGCACATCAT GTACACACTC CCGTCCACGA CCGACCCCCG CTGTTTTATT TTAATAGCTA 7350 CAAAGCAGGG AAATCCCTGC TAAAATGTCC TTTAACAAAC TGGTTAAACA AACGGGTCCA TCCGCACGGT 7420 GGACAGTTCC TCACAGTGAA GAGGAACATG CCGTTTATAA AGCCTGCAGG CATCTCAAGG GAATTACGCT 7490 GAGTCAAAAC TGCCACCTCC ATGGGATACG TACGCAACAT GCTCAAAAAG AAAGAATTTC ACCCCATGGC 7560 AGGGGAGTGG TTAGGGGGGT TAAGGACGGT GGGGGCGGCA GCTGGGGGCT ACTGCACGCA CCTTTTACTA 7630 AAGCCAGTTT CCTGGTTCTG ATGGTATTGG CTCAGTTATG GGAGACTAAC CATAGGGGAG TGGGGATGGG 7700 GGAACCCGGA GGCTGTGCCA TCTTTGCCAT GCCCGAGTGT CCTGGGCAGG ATAATGCTCT AGAGATGCCC 7770 ACGTCCTGAT TCCCCCAAAC CTGTGGACAG AACCCGCCCG GCCCCAGGGC CTTTGCAGGT GTGATCTCCG 7840 TGAGGACCCT GAGGTCTGGG ATCCTTCGGG ACTACCTGCA GGCCCGAAAA GTAATCCAGG GGTTCTGGGA 7910 AGAGGCGGGC AGGAGGGTCA GAGGGGGGCA GCCTCAGGAC GATGGAGGCA GTCAGTCTGA GGCTGAAAAG 7980 GGAGGGAGGG CCTCGAGCCC AGGCCTGCAA GCGCCTCCAG AAGCTGGAAA AAGCGGGGAA GGGACCCTCC 8050 ACGGAGCCTG CAGCAGGAAG GCACGGCTGG CCCTTAGCCC ACCAGGGCCC ATCGTGGACC TCCGGCCTCC 8120 GTGCCATAGG AGGGCACTCG CGCTGCCCTT CTAGCATGAA GTGTGTGGGG ATTTGCAGAA GCAACAGGAA 8190 ACCCATGCAC TGTGAATCTA GGATTATTTC AAAACAAAGG TTTACAGAAA CATCCAAGGA CAGGGCTGAA 8260 GTGCCTCCGG GCAAGGGCAG GGCAGGCACG AGTGATTTTA TTTAGCTATT TTATTTTATT TACTTACTTT 8330 CTGAGACAGA GTTATGCTCT TGTTGCCCAG GCTGGAGTGC AGCGGCATGA TCTTGGCTCA CTGCAACCTC 8400 CGTCTCCTGG GTTCAAGCAA TTCTCGTGCC TCAGCCTCCC AAGTAGCTGG GATTTCAGGC GTGCACCACC 8470 ACACCCGGCT AATTTTGTAT TTTTAGTAGA GATGGGCTTT CACCATGTTG GTCAAGCTGA TCTCAAAATC 8540 CTGACCTCAG GTGATCCGCC CACCTCAGCC TCCCAAAGTG CTGGGATTAC AGGCATGAGC CACTGCACCT 8610 GGCCTATTTA ACCATTTTAA AACTTCCCTG GGCTCAAGTC ACACCCACTG GTAAGGAGTT CATGGAGTTC 8680 AATTTCCCCT TTACTCAGGA GTTACCCTCC TTTGATATTT TCTGTAATTC TTCGTAGACT GGGGATACAC 8750 CGTCTCTTGA CATATTCACA GTTTCTGTGA CCACCTGTTA TCCCATGGGA CCCACTGCAG GGGCAGCTGG 8820 GAGGCTGCAG GCTTCAGGTC CCAGTGGGGT TGCCATCTCC CAGTAGAAAC CTGATGTAGA ATCAGGGCGC 8890 AAGTGTGGAC ACTGTCCTGA ATCTCAATGT CTCAGTGTGT GCTGAAACAT GTAGAAATTA AAGTCCATCC 8960 CTCCTACTCT ACTGGGATTG AGCCCCTTCC CTATCCCCCC CCAGGGGCAG AGGAGTTCCT CTCACTCCTG 9030 TGGAGGAAGG AATGATACTT TGTTATTTTT CACTGCTGGT ACTGAATCCA CTGTTTCATT TGTTGGTTTG 9100 TTTGTTTTGT TTTGAGAGGC GGTTTCACTC TTGTTGCTCA GGCTGGAGGG AGTGCAATGG CGCGATCTTG 9170 GCTTACTGCA GCCTCTGCCT CCCAGGTTCA AGTGATTCTC CTGCTTCCGC CTCCCATTTG GCTGGGATTA 9240 CAGGCACCCG CCACCATGCC CAGCTAATTT TTTGTATTTT TAGTAGAGAC GGGGGTGGGT GGGGTTCACC 9310 ATGTTGGCCA GGCTGGTCTC GAACTTCTGA CCTCAGATGA TCCACCTGCC TCTGCCTCCT AAAGTGCTGG 9380 GATTACAGGT GTGAGCCACC ATGCCCAGCT CAGAATTTAC TCTGTTTAGA AACATCTGGG TCTGAGGTAG 9450 GAAGCTCACC CCACTCAAGT GTTGTGGTGT TTTAAGCCAA TGATAGAATT TTTTTATTGT TGTTAGAACA 9520 CTCTTGATGT TTTACACTGT GATGACTAAG ACATCATCAG CTTTTCAAAG ACACACTAAC TGCACCCATA 9590 ATACTGGGGT GTCTTCTGGG TATCAGCAAT CTTCATTGAA TGCCGGGAGG CGTTTCCTCG CCATGCACAT 9660 GGTGTTAATT ACTCCAGCAT AATCTTCTGC TTCCATTTCT TCTCTTCCCT CTTTTAAAAT TGTGTTTTCT 9730 ATGTTGGCTT CTCTGCAGAG AACCAGTGTA AGCTACAACT TAACTTTTGT TGGAACAAAT TTTCCAAACC 9800 GCCCCTTTGC CCTAGTGGCA GAGACAATTC ACAAACACAG CCCTTTAAAA AGGCTTAGGG ATCACTAAGG 9870 GGATTTCTAG AAGAGCGACC TGTAATCCTA AGTATTTACA ACACGAGGCT AACCTCCAGC GAGCGTGACA 9940 GCCCAGGGAG GGTGCGAGGC CTGTTCAAAT GCTAGCTCCA TAAATAAAGC AATTTCCTCC GGCAGTTTCT 10010 GAAAGTAGGA AAGGTTACAT TTAAGGTTGC GTTTGTTAGC ATTTCAGTGT TTGCCGACCT CAGCTACAGC 10080 ATCCCTGCAA GGCCTCGGGA GACCCAGAAG TTTCTCGCCC CCTTAGATCC AAACTTGAGC AACCCGGAGT 10150 CTGGATTCCT GGGAAGTCCT CAGCTGTCCT GCGGTTGTGC CGGGGCCCCA GGTCTGGAGG GGACCAGTGG 10220 CCGTGTGGCT TCTACTGCTG GGCTGGAAGT CGGGCCTCCT AGCTCTGCAG TCCGAGGCTT GGAGCCAGGT 10290 GCCTGGACCC CGAGGCTGCC CTCCACCCTG TGCGGGCGGG ATGTGACCAG ATGTTGGCCT CATCTGCCAC 10360 ACAGAGTGCC GGGGCCCAGG GTCAAGGCCG TTGTGGCTGG TGTGAGGCGC CCGGTGCGCG GCCAGGAGGA 10430 GCGCCTGGCT CCATTTCCCA CCCTTTCTCG ACGGGACCGC CCCGGTGGGT GATTAACAGA TTTGGGGTGG 10500 TTTGCTCATG GTGGGGACCC CTCGCCGCCT GAGAACCTGC AAAGAGAAAT GACGGGCCTG TGTCAAGGAG 10570 CCCAAGTCGC GGGGAAGTGT TGCAGGGAGG CACTCCGGGA GGTCCCGCGT GCCCGTCCAG CGAGCAATGC 10640 GTCCTCGGGT TCGTCCCCAG CCGCCTCTAC GCCCCTCCGT CCTCCCCTTC ACGTCCGGCA TTCGTGGTGC 10710 CCGGAGCCCG ACGCCCCGCG TCCGGACCTG CAGGCAGCCC TGGGTCTCCG GATCAGGCCA GCGGCCAAAG 10780 GGTCGCCGCA CGCACCTGTT CCCAGGGCCT CCACATCATG GCCCCTCCCT CGGGTTACCC CACAGCCTAG 10850 GCCGATTCGA CCTCTCTCCC CTGGGGCCCT CGCTGGCGTC CCTGCACCCT GGGAGCGCGA GCGGCGCGCG 10920 GGCGGGGAAG CGCGGCCCAG ACCCCCGGGT CCGCCCGGAG CAGCTGCGCT GTCGGGGCCA GGCCGGGCTC 10990 CCAGTGGATT CGCGGGCACA GACGCCCAGG ACCGCGCTCC CCACGTGGCG GAGGGACTGG GGACCCGGGC 11060 ACCCGTCCTG CCCCTTCACC TTCCAGCTCC GCCTCCTCCG CGCGGACCCC GCCCCGTCCC GACCCCTCCC 11130 GGGTCCCCGG CCCAGCCCCC TCCGGGCCCT CCCAGCCCCT CCCCTTCCTT TCCGCGGCCC CGCCCTCTCC 11200 TCGCGGCGCG AGTTTCAGGC AGCGCTGCGT CCTGCTGCGC ACGTGGGAAG CCCTGGCCCC GGCCACCCCC 11270 GCGATGCCGC GCGCTCCCCG CTGCCGAGCC GTGCGCTCCC TGCTGCGCAG CCACTACCGC GAGGTGCTGC 11340 CGCTGGCCAC GTTCGTGCGG CGCCTGGGGC CCCACGGCTG GCGGCTGGTG CAGCGCGGGG ACCCGGCGGC 11410 TTTCCGCGCG CTGGTGGCCC AGTGCCTGGT GTGCGTGCCC TGGGACGCAC GGCCGCCCCC CGCCGCCCCC 11460 TCCTTCCGCC AGGTGGGCCT CCCCGGGGTC GGCGTCCGGC TGGGGTTGAG GGCGGCCGGG GGGAACCAGC 11550 GACATGCGGA GAGCAGCGCA GGCGACTCAG GGCGCTTCCC CCGCAGGTGT CCTGCCTGAA GGAGCTGGTG 11620 GCCCGAGTGC TGCAGAGGCT GTGCGAGCGC GGCGCGAAGA ACGTGCTGGC CTTCGGCTTC GCGCTGCTGG 11690 ACGGGGCCCG CGGGGGCCCC CCCGAGGCCT TCACCACCAG CGTGCGCAGC TACCTGCCCA ACACGGTGAC 11760 CGACGCACTG CGGGGGAGCG GGGCGTGGGG GCTGCTGCTG CGCCGCGTGG GCGACGACGT GCTGGTTCAC 11830 CTGCTGGCAC GCTGCGCGCT CTTTGTGCTG GTGGCTCCCA GCTGCGCCTA CCAGGTGTGC GGGCCGCCGC 11900 TGTACCAGCT CGGCGCTGCC ACTCAGGCCC GGCCCCCGCC ACACGCTAGT GGACCCCGAA GGCGTCTGGG 11970 ATGCGAACGG GCCTGGAACC ATAGCGTCAG GGAGGCCGGG GTCCCCCTGG GCCTGCCAGC CCCGGGTGCG 12040 AGGAGGCGCG GGGGCAGTGC CAGCCGAAGT CTGCCGTTGC CCAAGAGGCC CAGGCGTGGC GCTGCCCCTG 12110 AGCCGGAGCG GACGCCCGTT GGGCAGGGGT CCTGGGCCCA CCCGGGCAGG ACGCGTGGAC CGAGTGACCG 12180 TGGTTTCTGT GTGGTGTCAC CTGCCAGACC CGCCGAAGAA GCCACCTCTT TGGAGGGTGC GCTCTCTGGC 12250 ACGCGCCACT CCCACCCATC CGTGGGCCGC CAGCACCACG CAGGCCCCCC ATCCACATCG CGGCCACCAC 12320 GTCCCTGGGA CACGCCTTGT CCCCCGGTGT ACGCCGAGAC CAAGCACTTC CTCTACTCCT CAGGCGACAA 12390 GGAGCAGCTG CGGCCCTCCT TCCTACTCAG CTCTCTGAGG CCCAGCCTGA CTGGCGCTCG GAGGCTCGTG 12460 GAGACCATCT TTCTGGGTTC CAGGCCCTGG ATGCCAGGGA CTCCCCGCAG GTTGCCCCGC CTGCCCCAGC 12530 GCTACTGGCA AATGCGGCCC CTGTTTCTGG AGCTGCTTGG GAACCACGCG CAGTGCCCCT ACGGGGTGCT 12600 CCTCAAGACG CACTGCCCGC TGCGAGCTGC GGTCACCCCA GCAGCCGGTG TCTGTGCCCG GGAGAAGCCC 12670 CAGGGCTCTG TGGCGGCCCC CGAGGAGGAG GACACAGACC CCCGTCGCCT GGTGCAGCTG CTCCGCCAGC 12740 ACAGCAGCCC CTGGCAGGTG TACGGCTTCG TGCGGGCCTG CCTGCGCCGG CTGGTGCCCC CAGGCCTCTG 12810 GGGCTCCAGG CACAACGAAC GCCGCTTCCT CAGGAACACC AAGAAGTTCA TCTCCCTGGG GAAGCATGCC 12880 AAGCTCTCGC TGCAGGAGCT GACGTGGAAG ATGAGCGTGC GGGACTGCGC TTGGCTGCGC AGGAGCCCAG 12950 GTGAGGAGGT GGTGGCCGTC GAGGGCCCAG GCCCCAGAGC TGAATGCAGT AGGGGCTCAG AAAAGGGGGC 13020 AGGCAGAGCC CTGGTCCTCC TGTCTCCATC GTCACGTGGG CACACGTGGC TTTTCGCTCA GGACGTCGAG 13090 TGGACACGGT GATCTCTGCC TCTGCTCTCC CTCCTGTCCA GTTTGCATAA ACTTACGAGG TTCACCTTCA 13160 CGTTTTGATG GACACGCGGT TTCCAGGCGC CGAGGCCAGA GCAGTGAACA GAGGAGGCTG GGCGCGGCAG 13230 TGGAGCCGGG TTGCCGGCAA TGGGGAGAAG TGTCTGGAAG CACAGACGCT CTGGCGAGGG TGCCTGCAGG 13300 TTACCTATAA TCCTCTTCGC AATTTCAAGG GTGGGAATGA GAGGTGGGGA CGAGAACCCC CTCTTCCTGG 13370 GGGTGGGAGG TAAGGGTTTT GCAGGTGCAC GTGGTCAGCC AATATGCAGG TTTGTGTTTA AGATTTAATT 13440 GTGTGTTGAC GGCCAGGTGC GGTGGCTCAC GCCGGTAATC CCAGCACTTT GGGAAGCTGA GGCAGGTGGA 13510 TCACCTGAGG TCAGGAGTTT GAGACCAGCC TGACCAACAT GGTGAAACCC TATCTGTACT AAAAATACAA 13580 AAATTAGCTG GGCATGGTGG TGTGTGCCTG TAATCCCAGC TACTTGGGAG GCTGAGGCAG GAGAATCACT 13650 TGAACCCAGG AGGCGGAGGC TGCAGTGAGC TGAGATTGTG CCATTGTACT CCAGCCTGGG CGACAAGAGT 13720 GAAACTCTGT CTTTAAAAAA AAAAAGTGTT CGTTGATTGT GCCAGGACAG GGTAGAGGGA GGGAGATAAG 13790 ACTGTTCTCC AGCACAGATC CTGGTCCCAT CTTTAGGTAT GAAGAGGGCC ACATGGGAGC AGAGGACAGC 13860 AGATGGCTCC ACCTGCTGAG GAAGGGACAG TGTTTGTGGG TGTTCAGGGG ATGGTGCTGC TGGGCCCTGC 13930 CGTGTCCCCA CCCTGTTTTT CTGGATTTGA TGTTGAGGAA CCTCCGCTCC AGCCCCCTTT TGGCTCCCAG 14000 TGCTCCCAGG CCCTACCGTG GCAGCTAGAA GAAGTCCCGA TTTCACCCCC TCCCCACAAA CTCCCAAGAC 14070 ATGTAAGACT TCCGGCCATG CAGACAAGGA GGGTGACCTT CTTGGGGCTC TTTTTTTTCT TTTTTTCTTT 14140 TTATGGTGGC AAAAGTCATA TAACATGAGA TTGGCACTCC TAACACCGTT TTCTGTGTAC AGTGCAGAAT 14210 TGCTAACTCG GCGGTGTTTA CAGCAGGTTG CTTGAAATGC TGCGTCTTGC GTGACTGGAA GTCCCTACCC 14280 ATCGAACGGC AGCTGCCTCA CACCTGCTGC GGCTCAGGTG GACCACGCCG AGTCAGATAA GCGTCATGCA 14350 ACCCAGTTTT GCTTTTTGTG CTCCAGCTTC CTTCGTTGAG GAGAGTTTGA GTTCTCTGAT CAGGACTCTG 14420 CCTGTCATTG CTGTTCTCTG ACTTCAGATG AGGTCACAAT CTGCCCCTGG CTTATGCAGG GAGTGAGGCG 14490 TGGTCCCCGG GTGTCCCTGT CACGTGCAGG GTGAGTGAGG CGTTGCCCCC AGGTGTCCCT GTCACGTGTA 14560 GGGTGAGTGA GGCGCGGCCC CCGGGTGTCC CTGTCCCGTG CAGCGTGATT GAGGTGTGGC CCCCGGGTGT 14630 CCCTGTCACG TGTAGGGTGA GTGAGGCGCC ATCCCCGGGT GTCCCTGTCA CGTGTAGGGT GAGTGAGGCG 14700 TGGTCCCCGG GTGTCCCTGT CCCGTGCAGG GTGAGTGAGG CACTGTCCCC GGGTGTCCCT GTCACGTGCA 14770 GGGTGAGTGA GGCGCGGTCC CCGGGTGTCC CTCTCAGGTG TAGGGTGAGT GAGGCGCGGC CCCAGGGTGT 14840 CCCTGTCACG TGTAGGGTGA GTGAGGCACC GTCCCTGGGT GTCCCTCCCA GGTATAGGGT GAGTGAGGCA 14910 CTGTCCCCGG GTGTCCCTGT CACGTGCAGG GTGAGTGAGG CGCGGCCCCC GGGTGTCCCT CTCAGGTGCA 14980 GGGTGAGTGA GGCGCTGTCC CTGGGTGTCC CTGTCTCGTG TAGGGTGAGT GAGGCTCTGT CCCCAGGTGT 15050 CCTTGGCGTT TGCTCACTTG AGCTTGCTCC TGAATGTTTG CTCTTTCTAT AGCCACAGCT GCGCCGGTTG 15120 CCCATTGCCT GGGTAGATGG TGCAGGCGCA GTGCTGGTCC CCAAGCCTAT CTTTTCTGAT GCTCGGCTCT 15190 TCTTGGTCAC CTCTCCGTTC CATTTTGCTA CGGGGACACG GGACTGCAGG CTCTCGCCTC CCGCGTGCCA 15260 GGCACTGCAG CCACAGCTTC AGGTCCGCTT GCCTCTGTTG GGCCTGGCTT GCTCACCACG TGCCCGCCAC 15330 ATGCATGCTG CCAATACTCC TCTCCCAGCT TGTCTCATGC CGAGGCTGGA CTCTGGGCTG CCTGTGTCTG 15400 CTGCCACGTG TTGCTGGAGA CATCCCAGAA AGGGTTCTCT GTGCCCTGAA GGAAAGCAAG TCACCCCAGC 15470 CCCCTCACTT GTCCTGTTTT CTCCCAAGCT GCCCCTCTGC TTGGCCCCCT TGGGTGGGTG GCAACGCTTG 15540 TCACCTTATT CTGGGCACCT GCCGCTCATT GCTTAGGCTG GGCTCTGCCT CCAGTCGCCC CCTCACATGG 15610 ATTGACGTCC AGCCACAGGT TGGAGTGTCT CTGTCTGTCT CCTGCTCTGA GACCCACGTG GAGGGCCGGT 15680 GTCTCCGCCA GCCTTCGTCA GACTTCCCTC TTGGGTCTTA GTTTTGAATT TCACTGATTT TTAGTTTAGT 15750 TTTCTATCTC TCCATTGTAT GCTTTTTCTT GGTTTATTCT TTCATTCCTT TTCTAGCTTC TTAGTTTAGT 15820 CATGCCTTTC CCTCTAAGTG CTGCCTTACC TGCACCCTGT GTTTTGATGT GAAGTAATCT CAACATCAGC 15890 CACTTTCAAG TGTTCTTAAA ATACTTCAAA GTGTTAATAC TTCTTTTAAG TATTCTTATT CTGTGATTTT 15960 TTTCTTTGTG CACGCTGTGT TTTGA

GTGA AATCATTTTG ATATCAGTGA CTTTTAAGTA TTCTTTAGCT 16030 TATTCTGTGA TTTCTTTGAG CACTGAGTTA TTTGAACACT GTTTATGTTC AAGATATGTA GAGTATCAAG 16100 ATACGTAGAG TATTTTAAGT TATCATTTTA TTATTGATTT CTAACTCAGT TGTGTAGTGG TCTGTATAAT 16170 ACCAATTATT TGAAGTTTGC GGAGCCTTGC TTTGTGATCT AGTGTGTGCA TGGTTTCCAG AACTGTCCAT 16240 TGTAAATTTG ACATCCTGTC AATAGTGGGC ATGCATGTTC ACTATATCCA GCTTATTAAG GTCCAGTGCA 16310 AAGCTTCTGT CTCCTTCTAG ATGCATGAAA TTCCAAGAAG GAGGCCATAG TCCCTCACCT GGGGGATGGG 16380 TCTGTTCATT TCTTCTCCTT TGGTAGCATT TATGTGAGGC ATTGTTAGGT GCATGCACGT GGTAGAATTT 16450 TTATCTTCCT GATGAGTGAA TCTTTTGGAG ACTTCTATGT CTCTAGTAAT CTAGTAATTC TTTTTTTAAA 16520 TTGCTCTTAG TACTGCCACA CTGGGCTTCT TTTGATTAGT ATTTTCCTGC TGTGTCTGTT TTCTGCCTTT 16590 AATTTATATA TATATATATA TTTTTTTTTT TTTTGAGACA GAGTCTTGGT CTGTCGCCCA GGGTGAGTGC 16660 AGTGGTGTGA TCACAGGTCA GTGTAACTTT TACCTTCTGG CCTGAGCCGT CCTCTCACCT CAGCCTCCTG 16730 AGTAGCTGGA ACTGCAGACA CGCACCGCTA CACCTGGCTA ATTTTTAAAT TTTTTCTGGA GACAGGGTCT 16800 TGCTGTGTTG CCCAGGCTGG TCTCAAACTC TTGGACTCAA GGGATCCATC TACCTCGGCT TCCCAAAGTG 16870 CTGAATTACA GGCATGAGCC ACCATGTCTG GCCTAATTTT CAACACTTTT ATATTCTTAT AGTGTGGGTA 16940 TGTCCTGTTA ACAGCATGTA GGTGAATTTC CAATCCAGTC TGACAGTCGT TGTTTAACTG GATAACCTCA 17010 TTTATTTTCA TTTTTTTGTC ACTAGAGACC CGCCTGGTGC ACTCTGATTC TCCACTTGCC TGTTGCATGT 17080 CCTCGTTCCC TTGTTTCTCA CCACCTCTTG GGTTGCCATG TGCGTTTCCT GCCGAGTGTG TGTTGATCCT 17150 CTCGTTGCCT CCTGGTCACT GGGCATTTGC TTTTATTTCT CTTTGCTTAG TGTTACCCCC TGATCTTTTT 17220 ATTGTCGTTG TTTGCTTTTG TTTATTGAGA CAGTCTCACT CTGTCACCCA GGCTGGAGTG TAATGGCACA 17290 ATCTCGGCTC ACTGCAACCT CTGCCTCCTC GGTTCAAGCA GTTCTCATTC CTCAACCTCA TGAGTAGCTG 17360 GGATTACAGG CGCCCACCAC CACGCCTGGC TAATTTTTGT ATTTTTAGTA GAGATAGGCT TTCACCATGT 17430 TGGCCAGGCT GGTCTCAAAC TCCTGACCTC AAGTGATCTG CCCGCCTTGG CCTCCCACAG TGCTGGGATT 17500 ACAGGTGCAA GCCACCGTGC CCGGCATACC TTGATCTTTT AAAATGAAGT CTGAAACATT GGTACCCTTG 17570 TCCTGAGCAA TAAGACCCTT AGTGTATTTT AGCTCTGGCC ACCCCCCAGC CTGTGTGCTG TTTTCCCTGC 17640 TGACTTAGTT CTATCTCAGG CATCTTGACA CCCCCACAAG CTAAGCATTA TTAATATTGT TTTCCGTGTT 17710 GAGTGTTTCT GTAGCTTTGC CCCCGCCCTG CTTTTCCTCC TTTGTTCCCC GTCTGTCTTC TGTCTCAGGC 17780 CCGCCGTCTG GGGTCCCCTT CCTTGTCCTT TGCGTGGTTC TTCTGTCTTG TTATTGCTGG TAAACCCCAG 17850 CTTTACCTGT GCTGGCCTCC ATGGCATCTA GCGACGTCCG GGGACCTCTG CTTATGATGC ACAGATGAAG 17920 ATGTGGAGAC TCACGAGGAG GGCGGTCATC TTGGCCCGTG AGTGTCTGGA GCACCACGTG GCCAGCGTTC 17990 CTTAGCCAGT GAGTGACAGC AACGTCCGCT CGGCCTGGGT TCAGCCTGGA AAACCCCAGG CATGTCGGGG 18060 TCTGGTGGCT CCGCGGTGTC GAGTTTGAAA TCGCGCAAAC CTGCGGTGTG GCGCCAGCTC TGACGGTGCT 18130 GCCTGGCGGG GGAGTGTCTG CTTCCTCCCT TCTGCTTGGG AACCAGGACA AAGGATGAGG CTCCGAGCCG 18200 TTGTCGCCCA ACAGGAGCAT GACGTGAGCC ATGTGGATAA TTTTAAAATT TCTAGGCTGG GCGCGGTGGC 18270 TCACGCCTGT AATCCCAGCA CTTTGGGAGG CCAAGGCGGG TGGATCACGA GGTCAGGAGG TCGAGACCAT 18340 CCTGGCCAAC ATGATGAAAC CCCATCTGTA CTAAAAACAC AAAAATTAGC TGGGCGTGGT GGCGGGTGCC 18410 TGTAATCCCA GCTACTCGGG AGGCTGAGGC AGGAGAATTG CTTGAACCTG GGAGTTGGAA GTTGCAGTGA 18480 GCCGACATTG CACCACTGCA CTCCAGCCTG GCAACACAGC GAGACTCTGT CTCAAAAAAA AAAAAAAAAA 18550 AAAAAAAAAA AATTCTAGTA GCCACATTAA AAAAGTAAAA AAGAAAAGGT GAAATTAATG TAATAATAGA 18620 TTTTACTGAA GCCCAGCATG TCCACACCTC ATCATTTTAG GGTGTTATTG GTGGGAGCAT CACTCACAGG 18690 ACATTTGACA TTTTTTGAGC TTTGTCTGCG GGATCCCGTG TGTAGGTCCC GTGCGTGGCC ATCTCGGCCT 18760 GGACCTGCTG GGCTTCCCAT GGCCATGGCT GTTGTACCAG ATGGTGCAGG TCCGGGATGA GGTCGCCAGG 18830 CCCTCAGTGA GCTGGATGTG CAGTGTCCGG ATGGTGCACG TCTGGGATGA GGTCGCCAGG CCCTGCTGTG 18900 AGCTGGATGT GTGGTGTCTG GATGGTGCAG GTCAGGGGTG AGGTCTCCAG GCCCTCGGTG AGCTGGAGGT 18970 ATGGAGTCCG GATGATGCAG GTCCGGGGTG AGGTCGCCAG GCCCTGCTGT GAGCTGGATG TGTGGTGTCT 19040 GGATGGTGCA GGTCAGGGGT GAGGTCTCCA GGCCCTCGGT AAGCTGGAGG TATGGAGTCC GGATGATGCA 19110 GGTCCGGGGT GAGGTCGCCA GGCCCTGCTG TGAGCTGGAT GTGTGGTGTC TGGATGGTGC AGGTCTGGGG 19180 TGAGGTCACC AGGCCCTGCG GTGAGCTGGG TGTGCGGTGT CTGGATGGTG CAGGTCTGGA GTGAGGTCGC 19250 CAGACGGTGC CAGACCATGC GGTGAGCTGG ATATGCGGTG TCCGGATGGT GCAGGTCTGG GGTGAGGTTG 19320 CCAGGCCCTG CTGTGAGTTG GATGTGGGGT GTCCGGATGC TGCAGGTCCG GTGTGAGGTC ACCAGGCCCT 19390 GCTGTGAGCT GGATGTGTGG TGTCTGGATG GTGCAGGTCT GGGGTGAAGG TCGCCAGGCC CCTGCTTGTG 19460 AGCTGGATGT GTGGTGTCTG GATGGTGCAG GTCTGGAGTG AGGTCGCCAG GCCCTCGGTG AGCTGGATGT 19530 GCAGTGTCCA GATGGTGCAG GTCCGGGGTG AGGTCGCCAG ACCCTGCGGT GAGCTGGATG TGCCGTGTCT 19600 GGATGGTGCA GGTCTGGAGT GAGGTCGCCA GGCCCTCGGT GAGCTGGATG TATGGAGTCC GGATGGTGCC 19670 GGTCCGGGGT GAGGTCGCCA GACCCTGCTG TGAGCTGGAT GTGCGGTGTC TGGATGGTAC AGGTCTGGAG 19740 TGAGGTCGCC AGACCCTGCT GTGAGCTGGA TATGCGGTGT CCGGATGGTG CAGGTCAGGG GTGAGGTCTC 19810 CAGGCCCTCG GTGAGCTGGA GGTATGGAGT CCGGATGATG CAGGTCCGGG GTGAGGTCGC CAGGCCCTGC 19880 TGTGAACTGG ATGTGCGGCG TCTGGATGGT GCAGGTCTGG GGTGTGGTCG CCAGGCCCTC GGTGAGCTGG 19950 AGGTATGGAG TCCGGATGAT GCAGGTCCGG GGTGAGGTCG CCAGGCCCTG CTGTGAGCTG GATGTGCGGC 20020 GTCTGGATGG TGCAGGTCTG GGGTGTGGTC GCCAGGCCCT CGGTGAGCTG GAGGTATGGA GTCCGGATGA 20090 TGCAGGTCCG GGGTGAGGTT GCCAGGCCCT GCTGTGAGCT GGATGTGCTG TATCCGGATG GTGCAGTCCG 20160 GGGTGAGGTC GCCAGCCCCT GCTGTGAGCT GGATGTGCTC TATCCGGATG GTGCAGGTCT GGGGTGAGGT 20230 CACCAGGCCC TGCGGTGAGC TGGTTGTGCG GTGTCCGGTT CCTGCAGGTC CGGGGTGAGT TCGCCAGGCC 20300 CTCGGTGAGC TGGATGTGCG GTGTCCCCGT GTCCGGATGG TGCAGGTCCA GGGTCAGGTC GCTAGGCCCT 20370 TGGTGGGCTG GATGTGCCGT GTCCGGATGG TGCAGGTCTG GGGTGAGGTC GCCAGGCCTT TGGTGAGCTG 20440 GATGTGCGGT GTCTGCATGG TGCAGGTCTG GGGTGAGGTC GCCAGGCCCT TGGTGGGCTG GATGTGTGGT 20510 GTCCGGATGC TGCAGGTCCG GCGTGAGGTC GCCAGGCCCT GCTGTGAGCT GGATGTGCGG TGTCTGGATG 20580 GTGCAGGTCC GGGGTGAGGT AGCCAAGGCC TTCGGTGAGC TGGATGTGGG GTGTCCGGAT GGTGCAGGTC 20650 CGGGGTGAGG TCGCCAGGCC CTGCGGTTAG CTGGATATGC GGTGTCCGGA TGGTGCAGGT CCGGGGTGAG 20720 GTCACCAGGC CCTGCGGTTA GCTGGATGTG CGGTGTCTGC ATGGTGCAGG TCCGGGGTGA GGTCGCCAGG 20790 CCCTGCTGTG AGCTGGATGT GCTGTATCCG GATGGTGCAG GTCCGGGGTG AGGTCGCCAG GCCCTGCAGT 20860 GAGCTGGATG TGCTGTATCC GGATGGTGCA GGTCTGGCGT GAGGTCGCCA GGCCCTGCGG TTAGCTGGAT 20930 ATGCGGTGTC GGATGGTGCA GGTCCGGGGT GAGGTCACCA GGCCCTGCGG TTAGCTGGAT GTGCGGTGTC 21000 CGGATGGTGC AGGTCTGGGG TGAGGTCGCC AGGCCCTGCT GTGAGCTGGA TGTGCTGTAT CCGGATGGTG 21070 CAGGTCCGGG GTGAGGTCGC CAGGCCCTGC GGTGAGCTGG ATGTGCTGTA TCCGGATGGT GCAGGTCTGG 21140 CGTGAGGTCG CCAGGCCCTG CGGTGAGCTG GATGTGCAGT GTACGGATGG TGCAGGTCCG GGGTGAGGTC 21210 GCCAGGCCCT GCGGTGGGCT GTATGTGTGT TGTCTGGATG GTGCAGGTCC GGGGTGAGTT CGCCAGGCCC 21280 TGCGGTGAGC TGGATGTGTG GTGTCTGGAT GCTGCAGGTC CGGGGTGAGT TCGCCAGGCC CTCGGTGAGC 21350 TGGATATGCG GTGTCCCCGT GTCCGAATGG TGCAGGTCCA GGGTGAGGTC GCCAGGCCCT TGGTGGGCTG 21420 GATGTGCCGT GTCCGGATGG TGCAGGTCTC GGGTGAGGTC GCCAGGCCCT TGGTGAGCTG GATGTGCGGT 21490 GTCCGGATCG TGCAGGTCCG GGGTGAGGTC ACCAGGCCCT CGGTGATCTG GATGTGGCAT GTCCTTCTCG 21560 TTTAAGGGGT TGGCTGTGTT CCGGCCGCAG AGCACCGTCT GCGTGAGGAG ATCCTGGCCA AGTTCCTGCA 21630 CTGGCTGATG AGTGTGTACG TCGTCGAGCT GCTCAGGTCT TTCTTTTATG TCACGGAGAC CACGTTTCAA 21700 AAGAACAGGC TCTTTTTCTA CCGGAAGAGT GTCTGGAGCA ACTTGCAAAG CATTGGAATC AGGTACTGTA 21770 TCCCCACGCC AGGCCTCTGC TTCTCGAAGT CCTGGAACAC CAGCCCGGCC TCAGCATGCG CCTGTCTCCA 21840 CTTCCCTGTG CTTCCCTGGC TGTGCAGCTC TGGGCTGGGA GCCAGGGGCC CCGTCACAGG CCTGGTCCAA 21910 GTGGATTCTG TGCAAGGCTC TGACTGCCTG GAGCTCACGT TCTCTTACTT GTAAAATCAG GAGTTTGTGC 21980 CAAGTGGTCT CTAGGGTTTG TAAAGCAGAA GGGATTTAAA TTAGATGGAA ACACTACCAC TAGCCTCCTT 22050 GCCTTTCCCT GGGATGTGGG TCTGATTCTC TCTCTCTTTT TTTTTTCTTT TTTGAGATGG AGTCTCACTC 22120 TGTTGCCCAG GCTGGAGTGC AGTGGCATAA TCTTGGCTCA CTGCAACCTC CACCTCCTGG GTTTAAGCGA 22190 TTCACCAGCC TCAGCCTCCT AAGTAGCTGG GATTACAGGC ACCTGCCACC ACGCCTCGCT AATTTTTGTA 22260 CTTTTAGGAG AGACGGGGTT TCACCATGTT CGCCAGGCTG GTCTCGAACT CATGACCTCA GGTGATCCAC 22330 CCACCTTGGC CTCCCAAAGT GCTGGGTTTA CAGGCTAAGC CACCGTGCCC AGCCCCCGAT TCTCTTTTAA 22400 TTCATGCTGT TCTGTATGAA TCTTCAATCT ATTGGATTTA GGTCATGAGA CGATAAAATC CCACCCACTT 22470 GGCGACTCAC TGCAGGGAGC ACCTGTGCAG GGAGCACCTG GGGATAGGAG AGTTCCACCA TGAGCTAACT 22540 TCTAGGTGGC TGCATTTGAA TGGCTGTGAG ATTTTGTCTG CAATGTTCGG CTGATGAGAG TGTGAGATTG 22610 TGACAGATTC AAGCTGGATT TGCATCAGTG AGGGACGGGA GCGCTGGTCT GGGAGATGCC AGCCTGGCTG 22680 AGCCCAGGCC ATGGTATTAG CTTCTCCGTG TCCCGCCCAG GCTGACTGTG GAGGGCTTTA GTCAGAAGAT 22750 CAGGGCTTCC CCAGCTCCCC TGCACACTCG AGTCCCTGGG GGGCCTTGTG ACACCCCATG CCCCAAATCA 22820 GCATGTCTGC AGAGGGAGCT GGCAGCAGAC CTCGTCAGAG GTAACACAGC CTCTGGGCTG GGGACCCCGA 22890 CGTGGTGCTG GGGCCATTTC CTTGCATCTG GGGGAGGGTC AGGGCTTTCC CTGTGGGAAC AAGTTAATAC 22960 ACAATGCACC TTACTTAGAC TTTACACGTA TTTAATGGTG TGCGACCCAA CATGGTCATT TGACCAGTAT 23030 TTTGGAAAGA ATTTAATTGG GGTGACCGGA AGGAGCAGAC AGACGTGGTG GTCCCCAAGA TGCTCCTTGT 23100 CACTACTGGG ACTGTTGTTC TGCCTGGGGG GCCTTGGAGG CCCCTCCTCC CTGGACAGGG TACCGTGCCT 23170 TTTCTACTCT GCTGGGCCTG CGGCCTGCGG TCAGGGCACC AGCTCCGGAG CACCCGCGGC CCCAGTGTCC 23240 ACGGAGTGCC AGGCTGTCAG CCACAGATGC CCAGGTCCAG GTGTGGCCGC TCCAGCCCCC GTGCCCCCAT 23310 GGGTGGTTTT GGGGGAAAAG GCCAAGGGCA GAGGTGTCAG GAGACTGGTG GGCTCATGAG AGCTGATTCT 23380 GCTCCTTGGC TGAGCTGCCC TGAGCAGCCT CTCCCGCCCT CTCCATCTGA AGGGATGTGG CTCTTTCTAC 23450 CTGGGGGTCC TGCCTGGGGC CAGCCTTGGG CTACCCCAGT GCCTGTACCA GAGGGACAGG CATCCTGTGT 23520 GGAGGGGCAT GGGTTCACGT GGCCCCAGAT GCAGCCTGGG ACCAGGCTCC CTGGTGCTGA TGGTGGGACA 23590 GTCACCCTGG GGGTTGACCG CCGGACTGGG CGTCCCCAGG GTTGACTATA GGACCAGGTG TCCAGGTGCC 23660 CTGCAAGTAG AGGGGCTCTC AGAGGCGTCT GGCTGGCATG GGTGGACGTG GCCCCGGGCA TGGCCTTCAG 23730 CGTGTGCTGC CGTGGGTGCC CTGAGCCCTC ACTGAGTCGG TGGGGGCTTG TGGCTTCCCG TGAGCTTCCC 23800 CCTAGTCTGT TGTCTGGCTG AGCAAGCCTC CTGAGGGGCT CTCTATTGCA GACAGCACTT GAAGAGGGTG 23870 CAGCTGCGGG AGCTGTCGGA AGCAGAGGTC AGGCAGCATC GGGAAGCCAG GCCCGCCCTG CTGACGTCCA 23940 GACTCCGCTT CATCCCCAAG CCTGACGGGC TGCGGCCGAT TGTGAACATG GACTACGTCG TGGGAGCCAG 24010 AACGTTCCGC AGAGAAAAGA GGGTGGCTGT GCTTTGGTTT AACTTCCTTT TTAAACAGAA GTGCGTTTGA 24080 GCCCCACATT TGGTATCAGC TTAGATGAAG GGCCCGGAGG AGGGGCCACG GGACACAGCC AGGGCCATGG 24150 CACGGCGCCA ACCCATTTGT GCGCACAGTG AGGTGGCCGA GGTGCCGGTG CCTCCAGAAA AGCAGCGTGG 24220 GGGTGTAGGG GGAGCTCCTG GGGCAGGGAC AGGCTCTGAG GACCACAAGA AGCAGCCGGG CCAGGGCCTG 24290 GATGCAGCAC GGCCCGAGGT CCTGGATCCG TGTCCTGCTG TGGTGCGCAG CCTCCGTGCG CTTCCGCTTA 24360 CGGGGCCCGG GGACCAGGCC ACGACTGCCA GGAGCCCACC GGGCTCTGAG GATCCTGGAC CTTGCCCCAC 24430 GGCTCCTGCA CCCCACCCCT GTGGCTGCGG TGGCTGCGGT GACCCCGTCA TCTGAGGAGA GTGTGGGGTG 24500 AGGTGGACAG AGGTGTGGCA TGAGGATCCC GTGTGCAACA CACATGCGGC CAGGAACCCG TTTCAAACAG 24570 GGTCTGAGGA AGCTGGGAGG GGTTCTAGGT CCCGGGTCTG GGTGGCTGGG GACACTGGGG AGGGGCTGCT 24640 TCTCCCCTGG GTCCCTATGG TGGGGTGGGC ACTTGGCCGG ATCCACTTTC CTGACTGTCT CCCATGCTGT 24710 CCCCGCCAGG CCGAGCGTCT CACCTCGAGG GTGAAGGCAC TGTTCAGCGT GCTCAACTAC GAGCGGGCGC 24780 GGCGCCCCGG CCTCCTGGGC GCCTCTGTGC TGGGCCTGGA CGATATCCAC AGGGCCTGGC GCACCTTCGT 24850 GCTGCGTGTG CGGGCCCAGG ACCCGCCGCC TGAGCTGTAC TTTGTCAAGG TGGGTGCCGG GGACCCCCGT 24920 GAGCAGCCCT GCTGGACCTT GGGAGTGGCT GCCTGATTGG CACCTCATGT TGGGTGGAGG AGGTACTCCT 24990 GGGTGGGCCG CAGGGAGTGC AGGTGACCCT GTCACTGTTG AGGACACACC TGGCACCTAG GGTGGAGGCC 25060 TTCAGCCTTT CCTGCAGCAC ATGGGGCCGA CTGTGCACCC TGACTGCCCG GGCTCCTATT CCCAAGGAGG 25130 GTCCCACTGG ATTCCAGTTT CCGTCAGAGA AGGAACCGCA ACGGCTCAGC CACCAGGCCC CGGTGCCTTG 25200 CACCCCAGTC CTGAGCCAGG GGTCTCCTGT CCTGAGGCTC AGAGAGGGGA CACAGCCCGC CCTGCCCTTG 25270 GGGTCTGGAG TGGTGGGGGT CAGAGAGAGA GTGGGGGACA CCGCCAGGCC AGGCCCTGAG GGCAGAGGTG 25340 ATGTCTGAGT TTCTGCGTGG CCACTGTCAG TCTCCTCGCC TCCACTCACA CAGGTGGATG TGACGGGCGC 25410 GTACGACACC ATCCCCCAGG ACAGGCTCAC GGAGGTCATC GCCAGCATCA TCAAACCCCA GAACACGTAC 25480 TGCGTGCGTC GGTATGCCGT GGTCCAGAAG GCCGCCCATG GGCACGTCCG CAAGGCCTTC AAGAGCCACG 25550 TAAGGTTCAC GTGTGATAGT CGTGTCCAGG ATGTGTGTCT CTGGGATATG AATGTGTCTA GAATGCAGTC 25620 GTGTCTGTGA TGCGTTTCTG TGGTGGAGGT ACTTCCATGA TTTACACATC TGTGATATGC GTGTGTGGCA 25690 CGTGTGTGTC GTGGTGCATG TATCTGTGGC GTGCATATTT GTGGTGTGTG TGTGTGTGGC ACGTGTGTGT 25760 CCATGGTGTG TGTGCCTGTG GTGTGCATGT GTGTGTGTCT GTGACACGTG CATGTTCATG CTGTGTGCTG 25830 CATGTCTGTG ATGTGCCTAT TTGTGGTGTG TGTGTGCATG TGTCCGTGAC ATATGCGTGT CTATGGCATG 25900 GGTGTGTGTG GCCCCTTGGC CTTACTCCTT CCTCCTCCAG GCATGGTCCG CACCATTGTC CTCACGCTCT 25970 CGGGTGCTGG TTTGGGGAGC TCCACATTCA GGGTCCTCAC TTCTAGCATG GGTGCCCCTG TCCTGTCACA 26040 GGGCTGGGCC TTGGAGACTG TAAGCCAGGT TTGAGAGGAG AGTAGGGATG CTGGTGGTAC CTTCCTGGAC 26110 CCCTGGCACC CCCAGGACCC CAGTCTGGCC TATGCCGGCT CCATGAGATA TAGGAAGGCT GATTCAGGCC 26180 TCGCTCCCCG GGACACACTC CTCCCAGAGC GGCCGGGGGC CTTGGGGCTC GGCAGGGGTG AAAGGGGCCC 26250 TGGGCTTGGG TTCCCACCCA GTGGTCATGA GCACGCTGGA GGGGTAAGCC CTCAAAGTCG TGCCAGGCCG 26320 GGGTGCAGAG GTGAAGAAGT ATCCCTGGAG CTTCGGTCTG GGGAGAGGCA CATGTGGAAA CCCACAAGGA 26390 CCTCTTTCTC TGACTTCTTG AGCT 26414

Contig 2: TGTGGGATTG GTTTTCATGT GTGGGATAGG TGGGGATCTG TGGGATTGGT TTTTATGAGT GGGGTAACAC 70 AGAGTTCAAG GCGAGCTTTC TTCCTGTAGT GGGTCTGCAG GTGCTCCAAC AGCTTTATTG AGGAGACCAT 140 ATCTTCCTTT GAACTATGGT CGGGTTTATA GTAAGTCAGG GGTGTGGAGG CCTCCCCTGG GCTCCCTGTT 210 CTGTTTCTTC CACTCTGGGG TCGTGTGGTG CCTGCTGTGG TGTGTGGCCG GTGGGCAGGG CTTCCAGGCC 280 TCCTTGTGTT CATTGGCCTG GATGTGGCCC TGGCTACGCT CCGTCCTTGG AATTCCCCTG CGAGTTGGAG 350 GCTTTCTTTC TTTCTTTTTT TCTTTCTTTT TTTTTTTTTT TGATAACAGA GTCTCGCTCT TTTTTGCCCA 420 GGCTGGAGTG GTTTGGCGTG ATCTTGGCTC ACTGCAACCT GTGCTTCCTG AGTTCAAGCA ATTCTCTTGC 490 CTCAGCCTCC CAAGTAGCTG GAATTATAGG CGCCCACCAC CATGCTGACT AATTTTTGTA ATTTTAGTAG 560 AGACGAGGTT TCTCCATGTT GGCCAGGCTG GTCTCGAACT CCTGACCTCA GGTGATCCTC CCACCTCGGC 630 CTCCCAAAGT GCTGGGATGA CAGGTGTGAA CCGCCGCGCC CGGCCGAGAC TCGCTTCCTG CAGCTTCCGT 700 GAGATCTGCA GCGATAGCTG CCTGCAGCCT TGGTGCTGAC AACCTCCGTT TTCCTTCTCC AGGTCTCGCT 770 AGGGGTCTTT CCATTTCATG ACTCTCTTCA CAGAAGAGTT TCACGTGTGC TGATTTCCCG GCTGTTTCCT 840 GCGTAATTGG TGTCTGCTGT TTATCGATGG CCTCCTTCCA TTTCCTTTAG GCTTTGTTTA TTGTTGTTTT 910 TCCGGCTCCT TGAAGGAAAA GTTTCGATTA TGGATGTTTG AACTTTCTTT TCTAAACAAG CATCTGAAGT 980 TGCCGTTTTC CCTCTAAAGC AGGGATCCCG AGGCCCCTGG CTGTGGAGTG GCACCGGTCT GGGGCCTGTT 1050 AGGAACCCGG CGCACAGCGG GAGGCTAGGT GGGGTGTGGG GAGCCAGCGT TCCCGCCTGA GCCCCGCCCC 1120 TCTCAGATCA GCAGTGGCAT GCGGTGCTCA GAGGCGCACA CACCCTACTG AGAACTGTGC GTGAGAGGGG 1190 TCTAGATTCT GTGCTCCTTA TGGGAATCTA ATGCCTGATG ATCTGAGGTG GAACCGTTTG CTCCCAAAAC 1260 CATCCCCTTC CCCACTGCTG TCCTGTGGAA AAATCGTCTT CCACGAAACC AGTCCCTGGT ACCACAATGG 1330 TTGGGGACCC TGTGCTAAAG ACCTGCTTCA GCAGCCTCTC GTCAGTGTTG ATATATTGGC TTTTCTGTGT 1400 TGAGTCCAGA ATAATTACGG ATTTCTGTGA TGCTTTCCGC CGACCTCAGA CCCATGGGCT ATTTGTGGGC 1470 GTGTTGCCTG CTCCTGGGTT GGGAAGGGTG CAGGCCCCAT GTACCTTCCT GTTACTGCCT TCCAGGTTGG 1540 TTCTCAGGGT TGAATCGTAC TCGATGTGGT TTTAGCCCAC GGCCCTGCCG CCAGCTCCTG GGGGCTGGGG 1610 AACATGCTGA AGCACAGAGT CACCGTGCGC GTCTTTTGAT GCCTCACAAG CTCGAGGCCT CCTGTGTCCG 1680 TGTTAGTGTG TGTCACGTGC CTGCTCACAT CCTGTCTTGG GGACGCAGGG GCTTAGCAGG TCCCGTAGTA 1750 AATGACAAGC GTCCTGGGGG AGTCTGCAGA ATAGGAGGTG GGGGTGCCGG TCTCTCTCCC GCGTCTTCAG 1820 ACTCTTCTCC TGCCTGTGCT GTGGCTGCAC CTGCATCCCT GCAATCCCTC CAGCACTGGG CTGGAGAGGC 1890 CCGGGAGCTC GAGTGCCACT TGTGCCACGT GACTGTGGAT GGCAGTCGGT CACGGGGGTC TGATGTGTGG 1960 TGACTGTGGA TGGCGGTTGG TCACAGGGGT CTGATGTGTG GTGACTGTGG ATGGCGGTCG TGGGGTCTGA 2030 TGTGGTGACT GTGGATGGCG GTCGTGGGGT CTGATGTGTG GTGACTGTGG ATGGCGGTCG TGGGGTCTGA 2100 TGTGGTGACT GTGGATGGCG GTCGTGGGGT CTGATGTGGT GACTGTGGAT GGCGGTCGTG GGGTCTGATG 2170 TGGTGACTGT GGATGGCAGT CGTGGGGTCT GATGTGTGGT GACTGTGGAT GGCGGTCGTG GGGTCTGATG 2240 TGGTGACTGT GGATGGCAGT CGTGGGGTCT GATGTGTGGT GACTGTGGAT GGCGGTCGTG GGGTCTGATG 2310 TGTGGTGACT GTGGATGGCG GTCGTGGGGT CTGATGTGTG GTGACTGTGG ATGGCGGTCG TGGGGTCTGA 2380 TGTGTGGTGA CTGTGGATGG CGGTCGTGGG GTCTGATGTG GTGACTGTGG ATGGCGGTCG TGGGGTCTGA 2450 TGTGTGGTGA CTGTGGATGG TGATCGGTCA CAGGGGTCTG ATGTGTGGTG ACTGTGGATG GCGGTCGTGG 2520 GGTCTGATGT GTGGTGACTG TGGATGGTGA TCGGTCACAG GGGTCTGATG TGTGGTGACT GTGGATGGCG 2590 GTCGTGGGGT CTGATGTGTG GTGACTGTGG ATGGCGGTTG GTCCCGGGGG TCTGATGTGT GGTGACTGTG 2660 GATGGCGATC GGTCACAGGG GTCTGATGTG TGGTGACTGT GGATGGCGGT CGTGGGGTCT GATGTGTGGT 2730 GACTGTGGAT GGCGGTCGTG GGGTCTGATG TGTGGTGACT GTGGATGGCG GTCGTGGGGT CTGATGTGGT 2800 GACTGTGGAT GGCGGTCGTG GGGTCTGATG TGGTGACTGT GGATGGCGGT CGTGGGGTCT GATGTGTGGT 2870 GACTGTGGAT GGCGGTTGGT CCCGGGGGTC TGATGTGTGG TGACTGTGGA TGGCGGTCGT GGGGTCTGAT 2940 GTGGTGACTG TGGATGGCAG TCGTGGGGTC TGATGTGTGG TGACTGTGGA TGGCGGTCGT GGGGTCTGAT 3010 GTGTGGTGAC TGTGGATGGC GGTCGTGGGG TCTGATGTGT GGTGACTGTG GATGGCGGTC GTGGGGTCTG 3080 ATGTGTGGTG ACTGTGGATG GCGGTCGTGG GGTCTGATGT GGTGACTGTG GATGGCGGTC GTGGGGTCTG 3150 ATGTGTGGTG ACTGTGGATG GTGATCGGTC ACAGGGGTCT GATGTGTGGT GACTGTGGAT GGCGGTCGTG 3220 GGGTCTGATG TGTGGTGACT GTGGATGGCG GTCGTGGGGT CTGATGTGGT GACTGTGGAT GGCGGTCGTG 3290 GGGTCTGATG TGTGGTGACT GTGGATGGCG GTCGTAGGGT CTGATGTGTG GTGACTGTGG ATGGCAGTCG 3360 GTCACAGGGG TCTGATGTGT GGTGACTGTG GATGGCGGTC GTGGGGTCTG ATGTGTGGTG ACTGTGGATG 3430 GCGGTCGTGG GGTCTGATGT GTGGTGACTG TGGATGGCGG TCGTGGGGTC TGATGTGTGG TGACTGTGGA 3500 TGGCGGTCGT GGGGTCTGAT GTGGTGACTG TGGATGGTGA TCGGTCACAG GGGTCTGATG TGTGGTAGCT 3570 GCAGGTGGAG TCCCAGGTGT GTCTGTAGCT ACTTTGCGTC CTCGGCCCCC CGGCCCCCGT TTCCCAAACA 3640 GAAGCTTCCC AGGCGCTCTC TGGGCTTCAT CCCGCCATCG GGCTTGGCCG CAGGTCCACA CGTCCTGATC 3710 GGAAGAAACA AGTGCCCAGC TCTGGCCGGG GCAGGCCACA TTTGTGGCTC ATGCCCTCTC CTCTGCCGGC 3780 AGGTCTCTAC CTTGACAGAC CTCCAGCCGT ACATGCGACA GTTCGTGGCT CACCTGCAGG AGACCAGCCC 3850 GCTGAGGGAT GCCGTCGTCA TCGAGCAGGT CTGGGCACTG CCCTGCAGGG TTGGGCACGG ACTCCCAGCA 3920 GTGGGTCCTC CCCTGGGCAA TCACTGGGCT CATGACCGGA CAGACTGTTG GCCCTGGGGG GCAGTGGGGG 3990 GAATGAGCTG TGATGGGGGC ATGATGAGCT GTGTGCCTTG GCGAAATCTG AGCTGGGCCA TGCCAGGCTG 4060 CGACAGCTGC TGCATTCAGG CACCTGCTCA CGTTTGACTG CGCGGCCTCT CTCCAGTTCC GCAGTGCCTT 4130 TGTTCATGAT TTGCTAAATG TCTTCTCTGC CAGTTTTGAT CTTGAGGCCA AAGGAAAGGT GTCCCCCTCC 4200 TTTAGGAGGG CAGGCCATGT TTGAGCCGTG TCCTGCCCAG CTGGCCCCTC AGTGCTGGGT CTGAGGCCAA 4270 AGGAAACGTG TCCCCCTTCT TAGGAGGACG GGCCGTGTTT GAGCCACGCC CCGCTGAGCG GGCCTCTCAG 4340 TGCTGGGTCT GTCCACGTGG CCCTGTGGCC CTTTGCAGAT GTGGTCTGTC CACGTGGCCC TGTGGCTCTT 4410 TGCAGATGCC TGTTAGCACT TGCTCGGCTC TAGGGGACAG TCGTGTCCAC CGCATGAGGC TCAGAGACCT 4480 CTGGGCGAAT TTCCTTGGCT CCCAGGGTGG GGGTGGAGGT GGCCTGGGCT GCTGGGACCC AGACCCTGTG 4550 CCCGGCAGCT GGGCAGCAAC TCCTGGATCA CATATGCCAT CCGGGCCACG GTGGGCTGTG TGGGTGTGAG 4620 CCCAGCTGGA CCCACAGGTG GCCCAGAGGA GACGTTCTGT GTCACACACT TCGCCTAAGC CCATGTGTGT 4690 CTGCAGAGAC TCGGCCCGGC CAGCCCACGA TGGCCCTGCA TTCCAGCCCA GCCCCGCACT TCATCACAAA 4760 CACTGACCCC AAAAGGGACG GAGGGTCTTG GCCACGTGGT CCTGCCTGTC TCAGCACCCA CCGGCTCACT 4830 CCCATGTGTC TCCCGTCTGC TTTCGCAGAG CTCCTCCCTG AATGAGGCCA GCAGTGGCCT CTTCGACGTC 4900 TTCCTACGCT TCATGTGCCA CCACGCCGTG CGCATCAGGG GCAAGTGAGT CAGGTGGCCA GGTGCCATTG 4970 CCCTGCGGGC GGCTGGGCGG GCTGGCAGGG CTTCTGCTCA CCTCTCTCCT GCCCCTTCCC CACTGNCCTT 5040 CTGCCCGGGG CCACCAGAGT CTCCTTTTCT GGCCCCCGCC CCCTCCGGCT CCTGGGCTGC AGGCTCCCGA 5110 GGCCCCGGAA ACATGGCTCG GCTTGCGGCA GCCGGAGCGG AGCAGGTGCC ACACGAGGCC TGGAAATGGC 5180 AAGCGGGGTG TGGAGTTGCT CCTGCGTGGA GGACGAGGGG CGGGGGGTGT GTCTGGGTCA GGTGTGCGCC 5250 GAGCGTTTGA GCCTGCAGCT TGTCAGCTCC AAGTTACTAC TGACGCTGGA CACCCGGCTC TCACACGCTT 5320 CTATCTCTCT CTCCCGATAC AAAAGGATTT TATCCGATTC TCATTCCTGT CCCTGTCGTG TGACCCCCGC 5390 GAGGGCGCGG GCTCTTCTCT CTGTGACTAG ATTTCCCATC TGGAAAGTGC GGGGTTGACC GTGTAGTTTG 5460 CTCCTCTCGG GGGGCCTGTG GTGGCCATGG CGCAGGCGGC CTGGGAGAGC TGCCGTCACA CAGCCACTGG 5530 GTGAGCCACA CTCACGGTGG TAGAGCCACA GTGCCTGGTG CCACATCACG TCCTCTGGAT TTTAAGTAAA 5600 ACCACACACC TCCCGGCAGG CATCTGCCTG CGACCCTGTG TGTGCCTGGG GAGAGTGGTA GCACGGAGGA 5670 AATTCGTGCA CACTCAACGT CATCAGCAAG GTCATCCGCA GTCAGGTGGA ACGTGGAGGC CTCTCTCTGG 5740 GATCGTCTCC AGCGGATAAA GGACTGTGCA CAGCTTCGGA AGCTTTTATT TAAAAATATA ACTATTAATT 5810 ATTGCATTAT AAGTAATCAC TAATGGTATC AGCAATTATA ATATTTATTA AAGTATAATT AGAAATATTA 5880 AGTAGTACAC ACGTTCTGGA AAAACACAAA TTGCACATGG CAGCAGAGTG AATTTTGGCC GAGGGACACG 5950 TGTGCACATG TGTGTAAGCC GCCCCCAGGC CCACAGAATT CGCTGACAAA GTCACCTCCC CAGAGAAGCC 6020 ACCACGGGCC TCCTTCGTGG TCGTGAATTT TATTAACATG GATCAAGTCA CGTACCGTCC ACGTGTGGCA 6090 GGGCTTTGGG GAATGTGAGG TGATGACTGC GTCCTCATGC CCTGACAGAC AGGAGGTGAC TGTGTCTGTC 6160 CTGTCCCTAG GACACGGACA GGCCCGAAGC TCTAGTCCCC ATCGTCGTCC AGTTTGGCCT CTGAATAAAA 6230 ACGTCTTCAA AACCTGTTGC CCCAAAAACT AAGAACAGAG AGAGTTTCCC ATCCCATGTG CTCACAGGGG 6300 CGTATCTGCT TGCGTTGACT CGCTGGGCTG GCCGGACTCC TAGAGTTGGT GCGTGTGCTT CTGTGCAAAA 6370 AGTGCAGTCC TCTTGCCCAT CACTGTGATA TCTGCACCAG CAAGGAAAGC CTCTTTTCTT TTCTTTCTTT 6440 TTTTTTTTTT GAGACGGAAC GTCACTGTTG TCTGCCTGGG CTTGAGTGCA GTGGCGCGAT CTCAACTCAC 6510 TGCAACCTCC GCCTCCCGGG TTCCAGCATT TCTCCTGCCT CAGCCTCCCG AGCAGCTGAG ATTACAGGCA 6580 CCCACCCCCT GCGCCTGGCT AATTTTTGTA TTTTTAGTAG AGAGGGGTTT TTGCCATGTT GGCCAGGCTG 6650 GTCTCCAACT CCTGACCTCA GGTGATCCAC CCACCTCGGC CTCCCAAAGT GCTGGGATTA CAGGTGTGAG 6720 CCATCACGCC CAGCCGGAAA GCCTCTTTTT AAGGTGACCA CCTATAGCGC TTCCCGAAAA TAACAGGTCT 6790 TGTTTTTGCA GTAGGCTGCA AGCGTCTCTT AGCAACAGGA GTGGCGTCCT GTGGGCTCTG GGGATGGCTG 6860 AGGCTCGCGT GGCAGCCATG CCTTCTGTGT GCACCTTTAG GTTCCACGGG GCTATTCTGC TCTCACTGTT 6930 TGTCTGAAAA CGCACCCTTG GCATCCTTGT TTGGAGAGTT TCTGCTTCTC GTTGGTCATG CTGAAACTAG 7000 GGGCAAGGTT GTATCCGTTG GCGCGCAGCG GCTACATGTA GGGTCATGAG TCTTTCACCG TGGACAAATT 7070 CCTTGAAAAA AAAAAAAGGA GTCCGGTTAA GCATTCATTC CGGGTCAAGT GTCTGGTTCT GTGAATAAAC 7140 TCTAAGATTT AAGAAACCTT AATGAAAGAA AACCTTGATG ATTCAGAGCA AGGATGTGGT CACACCTGTG 7210 GCTGGATCTG TTTCAGCCGC CCCAGTGCAT GGTGAGAGTG GGGAGCAGGG ATTGTTTGTT CAGAGGTCTC 7280 ATCTGGTATG TTTCTGAGGT GTTTGCCGGC TGAATGGTAG ACGTGTCGTT TGTGTGTATG AGGTTCTGTG 7350 TCTGTGTGTG GCTCGGTTTG AGTGTACGCA TGTCCAGCAC ATGCCCTGCC CGTCTCTCAC CTGTGTCTTC 7420 CCGCCCCAGG TCCTACGTCC AGTGCCAGGG GATCCCGCAG GGCTCCATCC TCTCCACGCT GCTCTGCAGC 7490 CTGTGCTACG GCGACATGGA GAACAAGCTG TTTGCGGGGA TTCGGCGGGA CGGGTGAGGC CTCCTCTTCC 7560 CCAGGGGGGC TTGGGTGGGG GTTGATTTGC TTTTGATGCA TTCAGTGTTA ATATTCCTGG TGCTCTGGAG 7630 ACCATGACTG CTCTGTCTTG AGGAACCAGA CAAGGTTGCA GCCCCTTCTT GGTATGAAGC CGCACGGGAG 7700 GGGTTGCACA GCCTGAGGAC TGCGGGCTCC ACGCAGGCTC TGTCCAGCGG CCATGTCCAG AGGCCTCAGG 7770 GCTCAGCAGG CGGGAGGGCC GCTGCCCTGC ATGATGAGCA TGTGAATTCA ACACCGAGGA AGCACACCAG 7840 CTTCTGTCAC GTCACCCAGG TTCCGTTAGG GTCCTTGGGG AGATGGGGCT GGTGCAGCCT GAGGCCCCAC 7910 ATCTCCCAGC AGGCCCTCGA CAGGTGGCCT GGACTGGGCG CCTCTTCAGC CCATTGCCCA TCCCACTTGC 7980 ATGGGGTCTA CACCCAAGGA CGCACACACC TAAATATCGT GCCAACCTAA TGTGGTTCAA CTCAGCTGGC 8050 TTTTATTGAC AGCAGTTACT TTTTTTTTTT TAATACTTTA AGTTCTAGGG TACATGTGCA CGACGTGCAG 8120 GTTAGTTACA TATGTATACA TGTGCCATGT TGGTGTGCTG CACCCATTAA CTCATCATTT ACATTAGGTA 8190 TATCTCCTAA TGCTATCCCT CCCCACTCCC CCCATCCCAT GACAGGCCCT GGTGTGTGAT GTTCCCCACC 8260 CTGTGTCCAA GTGTTCTCAT TGTTCAGTTC CCACCTGTGA GTGAGAACAT GTGGTGTTTG GTTTTCTTTC 8330 CTTGCAATAG TTTGCTCAGA GTGATGGTTT CCAGCTTCGT CCATGTCCCT ACAAAGGACA TGAACTCATC 8400 CTTTTTTATG ACTGCATAGT ATTCCGTGGT GTATATGTGC CACATTTTCT TAATCCAGTC TATCATCGAT 8470 GGACATTTGG GTTGGTTGCA AGTCTTTGCT ACTGTGAATA GTGCCGCAAT AAACATACGT GTGCATGTGT 8540 CTTTATAGCA GCATGATTTA TAATCCTTTG GGTATATACC CAGTAATGGG ATGGCTGGGT CAAATGGTAT 8610 TTCTAGTTCT AGATCCTTGA GGAATCACCA CACTGTCTTC CACAATGGTT GAACTAGTTT ACACTCCCAC 8680 CAACAGTGTA AAAGTGTTCT GGTGCTGGAG AGGATGTGGA CAGCAGTTAT TTTTTTATGA AAATAGTATC 8750 ACTGAACAAG CAGACAGTTA GTGAAGGATG CGTCAGGAAG CCTGCAGGCC ACACAGCCAT TTCTCTCGAA 8820 GACTCCGGGT TTTTCCTGTG CATCTTTTGA AACTCTAGCT CCAATTATAG CATGTACAGT GGATCAAGGT 8890 TCTTCTTCAT TAAGGTTCAA GTTCTAGATT GAAATAAGTT TATGTAACAG AAACAAAAAT TTCTTGTACA 8960 CACAACTTGC TCTGGGATTT GGAGGAAAGT GTCCTCGAGC TGGCGGCACA CTGGTCAGCC CTCTGGGACA 9030 GGATACCTCT GGCCCATGGT CATGGGGCGC TGGGCTTGGG CCTGAGGGTC ACACAGTGCA CCATGCCCAG 9100 CTTCCTGTGG ATAGGATCTG GGTCTCGGAT CATGCTGAGG ACCACAGCTG CCATGCTGGT AAAGGGCACC 9170 ACGTCGCTCA GAGGGGGCGA GGTTCCCAGC CCCAGCTTTC TTACCGTCTT CAGTTATTTT TCCCTAAGAG 9240 TCTGAGAAGT GGGGCCGCGC CTGATGGCCT TCGTTCGTCT TCAGCTGGCA CAGAATTGCA CAAGCTGATG 9310 GTAAACACTG AGTACTTATA ATGAATGAGG AATTGCTGTA GCAGTTAACT GTAGAGAGCT CGTCTGTTGG 9380 AAAGAAATTT AAGTTTTTCA TTTAACCGCT TTGGAGAATG TTACTTTATT TATGGCTGTG TAAATTGTTT 9450 GACATTCAGT CCCTCGTAGA CAGATACTAC GTAAAAAGTG TAAAGTTAAC CTTGCTGTGT ATTTTCCCTT 9520 ATTTTAGGCT GCTCCTGCGT TTGGTGGATG ATTTCTTGTT GGTGACACCT CACCTCACCC ACGCGAAAAC 9530 CTTCCTCAGG TGAGGCCCGT GCCGTGTGTC TGTGGGGACC TCCACAGCCT GTGGGCTTTG CAGTTGAGCC 9650 CCCCGTGTCC TGCCCCTGGC ACCGCAGCGT TGTCTCTGCC AAGTCCTCTC TCTCTGCCGG TGCTGGATCC 9730 GCAAGAGCAG AGGCGCTTGG CCGTGCACCC AGGCCTGGGG GCGCAGGGGC ACCTTCGGGA GGGAGTGGGT 9800 ACCGTGCAGG CCCTGGTCCT GCACACACGC ACCCAGGTTA CACACGTGGT GAGTGCAGGC GGTGACCTGG 9870 CTCCTGCTGC TCTTTGGAAA GTCAAGAGTG GCGGCTCCTG GGGCCCCAGT GAGACCCCCA GGAGCTGTGC 9940 ACAGGGCCTG CAGGGCCGAG GCGGCAGCCT CCTCCCCAGG GTGCACCTGA GCCTGCGGAG AGCAGGAGCT 10010 GCTGAGTGAG CTGGCCCACA GCGTTCGCTG CGGTCACGTT CCTGCGTGGG GTTGTTTGGG ATCGGTGGGA 10080 GAATTTGGAT TTGCTGAGTG CTGCTGTCTT GAACCACGGA GATGGCTAGG AGTGGGTTTC AGAGTTGATT 10150 TTTGTGAATC AAACTAAAAT CAGGCACAGG GGACCTGGCC TCAGCACAGG GGATTGTCCA ATGTGGTCCC 10220 CCTCAAGGGC GCCCCACAGA GCCGGTGGGC TTGTTTTAAA GTGCGATTTG ACGAGGGACG AGAAACCTTG 10290 AAAGCTGTAA AGGGAACCCT CAGAAAATGT GGCCGCCAGG GGTGGTTTCA GGTGCTTTGC TGGGCTGTGT 10360 TTGTGAAAAC CCATTTGGAC CCGCCCTGCA AGTCCACCCT CCAGGTCCAC CCTCCAGGGC CGCCCTGGGC 10430 TGGGGGTATG CCTGGCGTTC CTTGTGCCGC AGCCCGGAGC ACAGCAGGCT GTGCACATTT AAATCCACTA 10500 AGATTCACTC GGGGGGAGCC CAGGTCCCAA GCAATTGAGG GCTCAGGAGT CCTGAGGCTG CTGAGGGGAC 10570 AGAGCAGACG GGGAACGCTG CTTCTGTGTG GCAACTTCCT GAGGGTGCTG GCCAGGGAGG TGGCTCAGAG 10640 TGTATGTTGG GGTCCCACCG GGGGCA

AAC TGTGTCTCTG ATGAGTCGGC AGCCATGTAA CAGGAAGGGG 10710 TGGCCACAGG GAGCTGGGAA TGCACCAGGG GAGCTGCGCA GCTGGCCGAG GTCCCAGGGC CAGGCCACAG 10780 GAAGGGCAGG GGGACGCCCG GGGCCACAGC AGAGGCCGCA CGAAGGGAAG GGGATGCCCA CGCCAGAGCA 10850 GAGGCTACCG GGCACAGGGG GGCTCCCTGA GCTGGGTGAG CGAGGCTCAT GACTCGGCCA GGGAACCTCC 10920 TTGACGTGAA GCTGACGACT GGTGTTGCCC AGCTCACAGC CCAGCCAGGT CCCGCGCCTG AGCAGGAACT 10990 CAGAACCCTC CCCTTTGTCT AAAGCACAGC AGATGCCTTC AGGGCATCTA GGAGAAAACA GGCAAAGTCG 11060 TTGAGAAACG TCTTAAAAGA AGGTGGGATG GTGGCAATTT CTTGTCCAGA TTTTAGTCTG CCCCGGACCA 11130 CAGATGAGTC TATAACGGGA TTGTGGTGTT GCCATGGGGA CACATGAGAT GGACCATCAC AGAGGCCACT 11200 GGGGCTGCAC CTCCCATCTG AGTCCTGGCT GTCCCGGGTC CAGGCCAGGT TCTTGCATGC TCACCTACCT 11270 GTCCTGCCCG GGAGACAGGG AAAGCACCCC GAAGTCTGGA GCAGGGCTGG GTCCAGGCTC CTCAGAGCTC 11340 CTGCCAGGCC CAGCACCCTG CTCCAAATCA CCACTTCTCT GGGGTTTTCC AAAGCATTTA ACAAGGGTGT 11410 CAGGTTACCT CCTGGGTGAC GGCCCCGCAT CCTGGGGCTG ACATTGCCCC TCTGCCTTAG GACCCTGGTC 11480 CGAGGTGTCC CTGAGTATGG CTGCGTGGTG AACTTGCGGA AGACAGTGGT GAACTTCCCT GTAGAAGACG 11550 AGGCCCTGGG TGGCACGGCT TTTGTTCAGA TGCCGGCCCA CGGCCTATTC CCCTGGTGCG GCCTGCTGCT 11620 GGATACCCGG ACCCTGGAGG TGCAGAGCGA CTACTCCAGG TGAGCGCACC TGGCCGGAAG TGGAGCCTGT 11690 GCCCGGCTGG GGCAGGTGCT GCTGCAGGGC CGTTGCGTCC ACCTCTGCTT CCGTGTGGGG CAGGCGACTG 11760 CCAATCCCAA AGGGTCAGAG GCCACAGGGT GCCCCTCGTC CCATCTGGGG CTGAGCAGAA ATGCATCTTT 11830 CTGTGGGAGT GAGGGTGCTC ACAACGGGAG CAGTTTTCTG TGCTATTTTG GTAAAAGGAA ATGGTGCACC 11900 AGACCTGGGT GCACTGAGGT GTCTTCAGAA AGCAGTCTGG ATCCGAACCC AAGACGCCCG GGCCCTGCTG 11970 GGCGTGAGTC TCTCAAACCC GAACACAGGG GCCCTGCTGG GCATGAGTCC CTCTGAACCC GAGACCCTGG 12040 GGCCCTGCTG GGCGTGAGTC TCTCCGAACC CAGAGACTTC AGGGCCCTTT TGGGCGTGAG TCTCTCCGCT 12110 GTGAGCCCCA CACTCCAAGG CTCATCCACA GTCTACAGGA TGCCATGAGT TCATGATCAC GTGTGACCCA 12180 TCAGGGGACA GGGCCATGGT GTGGGGGGGG TCTCTACAAA ATTCTGGGGT CTTGTTTCCC CAGAGCCCGA 12250 GAGCTCAAGG CCCCGTCTCA GGCTCAGACA CAAATGAATT GAAGATGGAC ACAGATGCAG AAATCTGTGC 12320 TGTTTCTTTT ATGAATAAAA AGTATCAACA TTCCAGGCAG GGCAAGGTGG CTCACACCTA TAATCCCAGC 12390 ACTTTGGGAG GCCGAGGTGG GTGGATCACT TGAGGCCAGG AGTTTGAGGC CAACCTAACC AACATAGTGA 12460 AATTCCATTT CTACTTAAAA AATACAAAAA TTAGCCTGGC CTGGTGGCAC ACGCCTGTAG TCCCCGCTAT 12530 GCGGGAGGCT GAGGCAGGAG AATCATTTGA ACCCAGGAGG CAGAGGTTGC AGTGAGCCGA GATCACACCA 12600 CTGCACTCCA GCCTGGGCAA CAGAGTGAGA CTTCATCTTA AAAAAAAAAA AAAAAGTATC AGCATTCCAA 12670 AACCATAGTG GACAGGTGTT TTTTTATTCT GTCCTTCGAT AATATTTACT GGTGCTGTGC TAGAGGCCGG 12740 AACTGGGGGT GCCTTCCTCT GAAAGGCACA CCTTCATGGG AAGAGAAATA AGTGGTGAAT GGTTGTTAAA 12810 CCAGAGGTTT AAACTGGGGT CCTGTCGTTC TGAGTTAACA GTCCAGATCT GGACTTTGCC TCTTTCCAGA 12880 ATGCTCCCTG GGGTTTGCTT CATGGGGGAG CAGCAGGTGT GGACACCCTC GTGATGGGGG AGCAGCAGGT 12950 GCAGACGCCC TCATGATGGG GGAGTGGCAG GTGCAGACAC CCTGGTGCAT GGTGCCCAGC ATGTCCCTGT 13020 TGCAGCTCCC TCCCCACAAG GATGCCGGTC TCCTGTGCTC CCCACAGTCC CTGCTTCCCT CTCACAGCCT 13090 TACCTGGTCC TGGCCTCCAC TGGCTTTGTC TGCATGATTT CCACATTTCC TGGGCTCCCA GCACCTCTTC 13160 GCCTCTCCCA GGCACCTCTG CAGTGCTGGC CATACCAGTC AGCTGTGAAC TGTCCACTGC TTATTTTGCT 13230 CCCCATGAAA TGTATTTTTT AGGACAGGCA CCCCTGGTTC CAGCCTCTGG CACAGCATCA GTGAATGTTA 13300 TTGAAGGACA AAGGACAGAC AAACAAATCA GGAAAATGGG TTCTCTCTAA ACACATTGCA AAGCCACAGA 13370 GGCTAGTGCA GGATGGGTGG GCATCAGGTC ATCAGATGTG GGTCCAATGC CAGAATATTC TGTGCTCCCA 13440 AAGGCCACTT GGTCAGAGTG TGTGCTTGCA GAGGTGGCTC TAAAAGCTCA GCAGTGGAGG CAGTGGTTCG 13510 CCATACTCAG GGTGAACTCA CATCCTCTGT GTCTGAAGTA TACAGCAGAG GCTTGAAGGG CATCTGGGAG 13580 AAGAAAACAG GCAAAATGAT TAAGAAAAGT GAAAAAGGAA AAGTGGTAAG ATGGGAATTT TCTTGTCCAG 13650 ATTTTAGTCT CCCAAACCAC AGCTCAGATG GTAGAATGTG GTCAGAACTG ATGGACAGAA CAATAGAACA 13720 AAACGGAAGC CCTATCTCTC AGAAACGTGT GTTAATGTGG TATGTGGCAC AGCTGATGGA AAAGAGAGTG 13790 TGTGTGTAAT TTTTTTTTCT GAGAAAACTG ACTGGAAGCA AATAAGTTGT GTCTTTACAG CATATACCAG 13860 AGCAGATTCT AGGTAGAAGA GGAGACACAT GCAAACAACA CCAGCAACAG AAATAAAACA AAAGACTCAA 13930 AGGGAAGGGA GGTGAACGTT CCCTGGTTTG GTGTTGGGGA AGGACACACA GGGAGGCGGA TGAAACCAGT 14000 GAGGCAACGG GCATTGCTTT CACTGCAGAG AAACTCAGCT TGCCTGAGCC ACAGTGAAAA TGGCCATTCC 14070 CTGGAGCGTT TGTGCACGTG ATTTATTTAA GGCGCCCTGT GAGGTCCTGC ACATTCATCC TCTCACTTTG 14140 TTCTCCTAAC CACCTGAGAG GTAGAGGAGG AAAGGCTCCA GGGGAGCAGC CGCCCTTGGT CACCCAGCTG 14210 GCAAAGGGCA TGCATGATTG CAGCCTGGCC TCCTGCTCCG GGGCCCTTGC TCTGCCCGAG GACCCCACAC 14280 AAGTCAGACC CATAGGCTCA GGGTGAGCCG GAGCCCAAGG TCGTGTTGGG GATGGCTGTG AAAGAAGAAA 14350 TGGACGTCTG ATGCACACTT GGGAAGGTCC TACCAGCAGC GTCAAAGAAA TGCATGTGAA ACTGACAGCG 14420 AGACCCATCC CTCAAAGAAA CGCACGTGAA ACTGATGGCG AGACCTGTCC CCATCCCTCA TGCTGGCTCC 14490 TTTTCTGGGC TTGCCAAGAG CCAGCATCAG GTTGAGGCAA GCTGGAAAGA CTTTTCTGGA AAGCAGCTTG 14560 TTTGCATGGA AGTCCTCACA ATGTCCTGTG TCTTCCCAGT AATTCCACTT CTGAAGTGAC CAGACATTAT 14630 CACGGGTCTT ATTTACCATT TCCAGTGTTC CAGGCAGGGG GACTTGCCAC AGCAAGTCAC GAACCTGCCC 14700 AAATACAGGG CTAAGGAGAT ATTATGCATC ACAAAACTTG CTCTGCCATT AAACATTTTT CAAAGAATTT 14770 TTGAAGAATG TTTAATGGCA CAAAACGTTT ATTTCAATGT AGCAGTGTTC AAAGCTGGAT GTAAAAGAAC 14840 ACACCCCAGG AGCCTGCCGT GAATGTCATG TGTGTTCATC TTTGGACATG GACATACATG GGCAGTGAGT 14910 GGTGGTGAGG CCCTGGAGGA CATCGGTGGG ATGCCTCCAT CCTGCCCCTC TGGAGACACC ATGTGTGCCA 14980 CGTGCACTCA CTGGAGCCCT GTTTAGCTGG TGCCACCTGG CTCTTCCATC CCTGAGATTC AAACACAGTG 15050 AGATTCCCCA CGCCCAACTC AGTGTTCTCC CACAAAAAAC CTGAGTCACA CCTGTGTTCA CTCGAGGGAC 15120 GCCCGGGAGC CAGGGCTCCA CAGTTTATTA TGTGTTTTTG GCTGAGTTAT GTGCAGATCT CATCAGGGCA 15190 GATGATGAGT GCACAAACAC GGCCGTGCGA GGTTTGGATA CACTCAACAT CACTAGCCAG GTCCTGGTGG 15260 AGTTTGGTCA TGCAGAGTCT GGATGGCATG TAGCATTTGG AGTCCATGGA GTGAGCACCC AGCCCCCTCG 15330 GGCTGCAGCG CATGCCCCAG GCAGGACAAG GAAGCGGGAG GAAGGCAGGA GGCTCTTTGG AGCAAGCTTT 15400 GCAGGAGGGG GCTGGGTGTG GGGCAGGCAC CTGTGTCTGA CATTCCCCCC TGTGTCTCAG CTATGCCCGG 15470 ACCTCCATCA GAGCCAGTCT CACCTTCAAC CGCGGCTTCA AGGCTGGGAG GAACATGCGT CGCAAACTCT 15540 TTGGGGTCTT GCGGCTGAAG TGTCACAGCC TGTTTCTGGA TTTGCAGGTG AGCAGGCTGA TGGTCAGCAC 15610 AGAGTTCAGA GTTCAGGAGG TGTGTGCGCA AGTATGTGTG TGTGTGTGTC CGCGCGTGCC TGCAAGGCTG 15680 ATGGTGACTG GCTGCACGTA AGAGTGCACA TGTACGCATA TACACGTGAG CACATACATG TCTGCATGTG 15750 TGTACATGAA GGCATGGCAG TGTGTGCACA GGTGTGCAAG GGCACAAGTG TGTGCACATG CGAATGCACA 15820 CCTGACATGC ATGTGTGTTC GTGCACAGTC GTGTGGGCAT TCACGTGAGG TGCATGCGTG TGGGTGTGCA 15990 GTGTGAGTAG CATGTGTGCA CATAACATGT ATTGAGGGGT CCTCGTGTTC ACCCCGCTAG GTCCTCAGCA 15960 CCAGTGCCAC TCCTTACAGG ATGAGACGGG GTCCCAGGCC TTGGTGGGCT GAGGCTCTGA AGCTGCAGCC 16030 CTGAGGGCAT TGTCCCATCT GGGCATCCGC GTCCACTCCC TCTCCTGTGG GCTTCTGTGT CCACTCCCCC 16100 TCTCCTGTGG GCATTTACAT CCACTCCACT CCCTCTCTCC TGTGGGCATC CGCGTCCACT CCCCCTCTCT 16170 GTGGGCATCT GCGTCCACCT CCCCTCTCTG TGGGCATTTG CGTCCACTCC CTCTCCTGGT TCCTTCCTGT 16240 CTTGGCCGAG CCTCGGGGGC AGGCAGATGA CACAGAGTCT TGACTCGCCC AGGGTGGTTC GCAGCTGCCG 16310 GGTGAGGGCC AGGCCGGATT TCACTGGGAA GAGGGATAGT TTCTTGTCAA AATGTTCCTC TTTCTTGTTC 16380 CATCTGAATG GATGATAAAG CAAAAAGTAA AAACTTAAAA TCCCAGAGAG GTTTCTACCG TTTCTCACTC 16450 TTTCTTGGCG ACTCTAGGTG AACAGCCTCC AGACGGTGTG CACCAACATC TACAAGATCC TCCTGCTCCA 16520 GGCGTACAGG TGAGCCGCCA CCAAGGGGTG CAGGCCCAGC CTCCAGGGAC CCTCCGCGCT CTGCTCACCT 16590 CTGACCCGGG GCTTCACCTT GGAACTCCTG GGTTTTAGGG GCAAGGAATG TCTTACGTTT TCAGTGGTGC 16660 TGCTGCCTGT GCACAGTTCT GTTCGCGTGG CTCTGTGCAA AGCACCTGTT CTCCATCTCT GGGTAGTGGT 16730 AGGAGCCGGT GTGGCCCCAG GTGTCCCCAC TGTGCCTGTG CACTGGCCGT GGGACGTCAT GGAGGCCATC 16800 CCAGGGCAGC AGGGGCATGG GGTAAAGAGA TGTTTATGGG GAGTCTTAGC AGAGGAGGCT GGGAAGGTGT 16870 CTGAACAGTA GATGGGAGAT CAGATGCCCG GAGGATTTGG GGTCTCAGCA AACAGGCCCG AGGTGGGTGC 16940 AGGTGAGGGT CGCTGGCCCC ACCCCCGGGA AGGTGCAGCA GAGCTGTGGC TCCCCACACA GCCCGGCCAG 17010 CACCTGTGCT CTGGGCATGG CTGTGCTCCT GGAACGTTCC CTGTCCTGGC TGGTCAGGGG GTGCCCCTGC 17080 CAAGAATCGA CAACTTTATC ACAGAGGGAA GGGCCAATCT CTGGAGGCCA CAGGGCCAGC TTCTGCCTGG 17150 AGTCAGGGCA GGTGGTGGCA CAAGCCTCGG GGCTGTACCA AAGGGCAGTC GGGCACCACA GGCCCGGGCC 17220 TCCACCTCAA CAGGCCTCCC GAGCCACTGG GAGCTGAATG CCAGGAGGCC CAAGCCCTCG CCCCATGAGG 17290 GCTGAGAAGG AGTGTGAGCA TTTGTGTTAC CCAGGGCCGA GGCTGCGCGA ATTACCGTGC ACACTTGATG 17360 TGAAATGAGG TCGTCGTCTA TCGTGGAAAC CCAGCAAGGG CTCACGGGAG AGTTTTCCAT TACAAGGTCG 17430 TACCATGAAA ATGGTTTTTA ACCCGAGTGC TTGCGCCTTC ATGCTCTGGC AGGGAGGGCA GAGCCACAGC 17500 TGCATGTTAC CGCCTTTGCA CCAGCTCCAG AGGCTTGGGA CCAGGCTGTC TCAGTTCCAG GGTCCGTCCG 17570 GCTCAGACCG CCCTCCTCTC TGCCTTCTCT CTCTGCCTCA AATCTTCCCT CGTTTGCATC TCCCTGACGC 17640 GTGCCTGGGC CCTCGTGCAA GCTGCTTGAC TCCTTTCCGG AAACCCTTGG GGTGTGCTGG ATACAGGTGC 17710 CACTGAGGAC TGGAGGTGTC TGACACTGTG GTTGACCCCA GGGTCCAGCT GGCGTGCTTG GGGCCTCCTT 17780 GGGCCATGAT GAGGTCAGAG GAGTTTTCCC AGGTGAAAAC TCCTGGGAAA CTCCCAGGGC CATGTGACCT 17850 GCCACCTGCT CCTCCCATAT TCAGCTCAGT CTTGTCCTCA TTTCCCCACC AGGGTCTCTA GCTCCGAGGA 17920 GCTCCCGTAG AGGGCCTGGG CTCAGGGCAG GGCGGCTGAG TTTCCCCACC CATGTGGGGA CCCTTGGGTA 17990 GTCGCTTGAT TGGGTAGCCC TGAGGAGGCC GAGATGCGAT GCGCCACGGG CCGTTTCCAA ACACAGAGTC 18060 AGGCACGTGG AAGGCCCAGG AATCCCCTTC CCTCGAGGCA GGAGTGGGAG AACGGAGAGC TGGGCCCCGA 18130 TTTCACGGCA GCCAGGCTGC AGTGGGCGAG GCTGTGGTGG TCCACGTGGC GCTGGGGGCG GGGTCTGATT 18200 CAAATCCGCT GGGGCTCGGC CTTCCTGGCC CGTGCTGGCC GCGCCTCCAC ACGGGCTTGG GGTGGACGCC 18270 CCGACCTCTA GCAGGTGGCT ATTTCTCCCT TTGGAAGAGA GCCCCTCACC CATGCTAGGT GTTTCCCTCC 18340 TGGGTCAGGA GCGTGGCCGT GTGGCAACCC CGGGACCTTA GGCTTATTTA TTTGTTTAAA AACATTCTGG 18410 GCCTGGCTTC CGTTGTTGCT AAATGGGGAA AAGACATCCC ACCTCAGCAG AGTTACTGAG AGGCTGAAAC 18480 CGGGGTGCTG GCTTGACTGG TGTGATCTCA GGTCATTCCA GAAGTGGCTC AGGAAGTCAG TGAGACCAGG 18550 TACATGGGGG GCTCAGGCAG TGGGTGAGAT GAGGTACACG GGGGGCTCAG GCAGTGGGTG AGGCCAGGTA 18620 CATGGGGGGC TCAGGCACTG GGTGAGATGA GGTACACGGG GGGCTCAGGC AGAGGGTCAG ACCAGGTACA 18690 CGGGGGCTCT GATCACACGC ACATATGAGC ACATGTGCAC ATGTGCTGTT TCATGGTAGC CAGGTCTGTG 18760 CACACCTGCC CCAAAGTCCC AGGAAGCTGA GAGGCCAAAG ATGGAGGCTG ACAGGGCTGG CGCGGTGGCT 18830 CACACCTGTA GTCCCAGCAC TTTGGGAGGC CGAGGCGAGA GGATCCCTTG AGCCCAGGAG TTTAAGACCA 18900 GCCTGAGCAA CATAGTAGAA CCCCATCTCT ATGAAAAATA AAAACAAAAA TTAGCTGAAC ATGGTGGTGT 18970 GCGCCTGTAG TTCCAATACT TGGGAGGCTG AAGTGGGAGG ATCACTTGAG CCCAGGAGGT GGAAGCTGCA 19040 GTGAGCTGAG ATTGCACCAC TGTACTGCAG CCTGGGTGAC AGAGTGAGAG CCCATCTCAA CAACAACAAA 19110 GAAGACTGAC AAATGCAGTT TCTTGGAAAG AAACATTTAG TAGGAACTTA ACCTACACAC AGAAGCCAAG 19180 TCGGTGTCTC GGTGTCAGTG AGATGAGATG ATGCGTCCTC ACACCATCAC CCCAGACCCA GGGTTTATGC 19250 ACCACAGGGG CGGGTGGCTC AGAAGGGATG CGCAGGACGT TGATATACGA TGACATCAAG GTTGTCTGAC 19320 GAAGGGCAGG ATTCATGATA AGTACCTGCT GGTACACAAG GAACAATGGA TAAACTGGAA ACCTTAGAGG 19390 CCTTCCCGGA ACAGGGGCTA ATCAGAAGCC AGCATGGGGG GCTGGCATCC AGGATGGAGC TGCTTCAGCC 19460 TCCACATGCG TGTTCATACA GATGGTGCAC AGAAACGCAC TGTACCTGTG CACACACAGA CACGCAGCTA 19530 CTCGCACACA CAAGCACACA CACAGACATG CATGCATGCA TCCGTGTGTG TGCACCTGTG CCCATGAGGA 19600 AACCCATGCA TGTGCATTCA TGCACGCACA CAGGCACCGG TGGGCCCATG CCCACACCCA CGAGCACCGT 19670 CTGATTAGGA GGCCTTTCCT CTGACGCTGT CCGCCATCCT CTCAGGTTTC ACGCATGTGT GCTGCAGCTC 19740 CCATTTCATC AGCAAGTTTG GAAGAACCCC ACATTTTTCC TGCGCGTCAT CTCTGACACG GCCTCCCTCT 19810 GCTACTCCAT CCTGAAAGCC AAGAACGCAG GTATGTGCAG GTGCCTGGCC TCAGTGGCAG CAGTGCCTGC 19880 CTGCTGGTGT TAGTGTGTCA GGAGACTGAG TGAATCTGGG CTTAGGAAGT TCTTACCCCT TTTCGCATCA 19950 GGAAGTGGTT TAACCCAACC ACTGTCAGGC TCGTCTGCCC GCCCTCTCGT GGGGTGAGCA GAGCACCTGA 20020 TGGAAGGGAC AGGACCTGTC TGGGAGCTGC CATCCTTCCC ACCTTGCTCT GCCTGGGGAA GCGCTGGGGG 20090 GCCTGGTCTC TCCTGTTTGC CCCATGGTGG GATTTGGGCG GCCTGGCCTC TCCTGTTTGC CCTGTGGTGG 20160 GATTGGGCTG TCTCCCGTCC ATGGCACTTA GGGCCCTTGT GCAAACCCAG GCCAAGGGCT TAGGAGGAGG 20230 CCAGCCCCAG GCTACCCCAC CCCTCTCAGG AGCAGAGGCC GCGTATCACC ACGACAGAGC CCCGCGCCGT 20300 CCTCTGCTTC CCAGTCACCG TCCTCTGCCC CTGGACACTT TGTCCAGCAT CAGGGAGGTT TCTGATCCGT 20370 CTGAAATTCA AGCCATGTCG AACCTGCGGT CCTGAGCTTA ACAGCTTCTA CTTTCTGTTC TTTCTGTGTT 20440 GTGGAAATTT CACCTGGAGA AGCCGAAGAA AACATTTCTG TCGCGACTCC TGCGGTGCTT GGGTCGGGAC 20510 AGCCAGAGAT GGAGCCACCC CGCAGACCGT CGGGTGTGGG CAGCTTTCCG GTCTCTCCTG GGAGGGGAGC 20580 TGGGCTGCGC CTGTGACTCC TCAGCCTCTG TTTTCCCCCA GGGATGTCGC TGGGGGCCAA GGCCGCCCCC 20650 GGCCCTCTGC CCTCCGAGGC CGTGCAGTGG CTGTGCCACC AAGCATTCCT GCTCAAGCTG ACTCGACACC 20720 GTGTCACCTA CGTGCCACTC CTGGGGTCAC TCAGGACAGG GAAGTGTGGG TGGACGCCAG TGCGGGCCCC 20790 ACCTGCCCAG GGGTCATCCT TGAACGCCCT GTGTGGGGCG AGCAGCCTCA GATGCTGCTG AAGTGCAGAC 20860 GCCCCCGGGC CTGACCCTGG GGGCCTGGAG CCACGCTGGC AGCCCTATGT GATTAAACGC TGGTGTCCCC 20930 AGGCCACGGA GCCTGGCAGG GTCCCCAACT TCTTGAACCC CTGCTTCCCA TCTCAGGGGC GATGGCTCCC 21000 CACGCTTGGG AGCCTTCTGA CCCCTGACCT GTGTCCTCTC ACAGCCTCTT CCCTGGCTGC TGCCCTGAGC 21070 TCCTGGGGTC CTGAGCAAGT TCTCTCCCCG CCCCGCCGCT CCAGCGTCAC TGGGCTGCCT GTCTGCTCGC 21140 CCCGGTGGAG GGGTGTCTGT CCCTTCACTG AGGTTCCCAC CAGCCAGGGC CACGAGGTGC AGGCCCTGCC 21210 TGCCCGGCCA CCCACACGTC CTAGGAGGGT TGGAGGATGC CACCTCTGGC CTCTTCTGGA ACGGAGTCTG 21280 ATTTTGGCCC CGCAGCCCAG ACGCAGCTGA GTCGGAAGCT CCCGGGGACG ACGCTGACTG CCCTGGAGGC 21350 CGTAGCCAAC CCGGCACTGC CCTCAGACTT CAAGACCATC CTGGACTGAT GGCCACCCGC CCACAGCCAG 21420 GCCGAGAGCA GACACCAGCA GCCCTGCCAC GCCGGGCTCT ACGTCCCAGG GAGGGAGGGG CGGCCCACAC 21490 CCAGGCCCGC ACCGCTGGGA GTCTGAGGCC TGAGTGAGTG TTTGGCCGAG GCCTGCATGT CCGGCTGAAG 21560 GCTGAGTGTC CGGCTGAGGC CTGAG

GAGT GTCCAGCCAA GGG

TGAGTG TCCAGCACAC CTGCCGTCTT 21630 CACTTCCCCA CAGGCTGGCG CTCGGCTCCA CCCCAGGGCC AGCTTTTCCT CACCAGGAGC CCGGCTTCCA 21700 CTCCCCACAT AGGAATAGTC CATCCCCAGA TTCGCCATTG TTCACCCCTC GCCCTGCCCT CCTTTGCCTT 21770 CCACCCCCAC CATCCAGGTG GAGACCCTGA GAAGGACCCT GGGAGCTCTG GGAATTTGGA GTGACCAAAG 21840 GTGTGCCCTG TACACAGGCG AGGACCCTGC ACCTCGATGG CGGTCCCTGT GGGTCAAATT CGGGGGAGGT 21910 GCTGTGGGAG TAAAATACTG AATATATGAG TTTTTCAGTT TTGAAAAAAA TCTCATGTTT GAATCCTAAT 21980 GTGCACTGCA TAGACACCAC TGTATGCAAT TACAGAAGCC TGTGAGTGAA CGGGGTGGTG GTCAGTGCGG 22050 GCCCATGGCC TGGCTGTGCA TTTACGGAAG TCTATGAGTG AATGGGGTTG TGGTCAGTGC GGGCCCATGG 22120 CCTGGCTGGG CCTGGGAGGT TTCTGATGCT GTGAGGCAGG AGGGGAAGGA GGGTAGGGGA TAGACAGTGG 22190 GAGCCCCCAC CCTGGAAGAC ATAACAGTAA GTCCAGGCCC GAAGGGCAGC AGGGATGCTG GGGGCCCAGC 22260 TTGGGCGGCG GGGATGATGG AGGGCCTGGC CAGGGTGGCA GGGATGATGG GGGCCCCAGC TGGGGTGGCA 22330 GGGGTGATGG GGGGGGCTGG TCTGGGTGGC GGGGAAGATG GGGAAGCCTG GCTGGGCCCC CTCCTCCCCT 22400 GCCTCCCACC TGCAGCCGTG GATCCGGATG TGCTTCCCTG GTGCACATCC TCTGGGCCAT CAGCTTTCAT 22470 GGAGGTGGGG GGCAGGGGCA TGACACCATC CTGTATAAAA TCCAGGATTC CTCCTCCTGA ACGCCCCAAC 22540 TCAGGTTGAA AGTCACATTC CGCCTCTGGC CATTCTCTTA AGAGTAGACC AGGATTCTGA TCTCTGAAGG 22610 GTGGGTAGGG TGGGGCAGTG GAGGGTGTGG ACACAGGAGG CTTCAGGGTG GGGCTGGTGA TGCTCTCTCA 22680 TCCTCTTATC ATCTCCCAGT CTCATCTCTC ATCCTCTTAT CATCTCCCAG TCTCATCTGT CTTCCTCTTA 22750 TCTCCCAGTC TCATCTGTCA TCCTCTTACC ATCTCCCAGT CTCATCTCTT ATCCTCTTAT CTCCTAGTCT 22820 CATCCAGACT TACCTCCCAG GGCGGGTGCC AGGCTCGCAG TGGAGCTGGA CATACGTCCT TCCTCAGGCA 22890 GAAGGAACTG GAAGGATTGC AGAGAACAGG AGGGGCGGCT CAGAGGGACG CAGTCTTGGG GTGAAGAAAC 22960 AGCCCCTCCT CAGAAGTTGG CTTGGGCCAC ACGAAACCGA GGGCCCTGCG TGAGTGGCTC CAGAGCCTTC 23030 CAGCAGGTCC CTGGTGGGGC CTTATGGTAT GGCCGGGTCC TACTGAGTGC ACCTTGGACA GGGCTTCTGG 23100 TTTGAGTGCA GCCCGGACGT GCCTGGTGTC GGGGTGGGGG CTTATGGCCA CTGGATATGG CGTCATTTAT 23170 TGCTGCTGCT TCAGAGAATG TCTGAGTGAC CGAGCCTAAT GTGTATGGTG GGCCCAAGTC CACAGACTGT 23240 GTCGTAAATG CACTCTGGTG CCTGGAGCCC CCGTATAGGA GCTGTGAGGA AGGAGGGGCT CTTGGCAGCC 23310 GGCCTGGGGG CGCCTTTGCC CTGCAAACTG GAAGGGAGCG GCCCCGGGCG CCGTGGGCGG ACGACCTCAA 23380 GTGAGAGGTT GGACAGAACA GGGCGGGGAC TTCCCAGGAG CAGAGGCCGC TGCTCAGGCA CACCTGGGTT 23450 TGAATCACAG ACCAACAGGT CAGGCCATTG TTCAGCTATC CATCTTCTAC AAAGCTCCAG ATTCCTGTTT 23520 CTCCGGGTGT TTTTTGTTGA AATTTTACTC AGGATTACTT ATATTTTTTG CTAAAGTATT AGACCCTTAA 23590 AAAAGGTATT TGCTTTGATA TGGCTTAACT CACTAAGCAC CTACTTTATT TGTCTGTTTT TATTTATTAT 23660 TATTATTATT ATTAGAGATG GTGTCTACTC TGTCACCCAG GTTGTTAGTG CAGTGGCACA GTCATGGCTC 23730 GCTGTAGCCG CAAACCCCCA GGCTCAAGTG ATCCTCCGGC CTCAGCTTCC CAGAGTGCTG GGATTACAGG 23800 TGTGAGCCAC TGCCCTTGCC TGGCACTTTT AAAAACCACT ATGTAAGGTC AGGTCCAGTG GCTTCCACAC 23870 CTGTCATCCC AGTAGTTTGG GAAGCCGAGG CAGAAGGATT GTCTGAGGCC AGGAGTTTGA GACCAGCATG 23940 GGTAACATAG GGAGACCCCA TCTCTACAAA AAATGCAAAA AGTTATCCGG GCGTGGGGTC CAGCATCTGT 24010 AGTCCCAGCT GCTCGGGAGG CTGAGTGGGA GGATCGCTTG AGCCCGGGAG GTCATGGCTG CAGTGAGCTG 24080 TGATTGTACC ATCGCACTCC AGCCTGGGCA ACAGAGTGAG ACCCTGTCTC AAAAAAAAAA AAAAAAAAAG 24150 AAGGAGAAGG AGAAGAGAAG AAGAAGGAAG AAGGAAAGAG AAGAAGAAGG AAGAAGGAAG AAAGAAGGAG 24220 AAGGAGGCCT GCTAGGTGCT AGGTAGACTG TCAAATCTCA GAGCAAAATG AAAATAACAA AGTTTTAAAG 24290 GGAAAGAAAA ACCCCAGCTC TTTGGACTTC CTTAGGCCTG AACTTCATCT CAAGCAGCTT CCTTCCACAG 24360 ACAAGCGTGT ATGGAGCGAG TGAGTTCAAA GCAGAAAGGG AGGAGAAGCA GGCAAGGGTG GAGGCTGTGG 24430 GTGACACCAG CCAGGACCCC TGAAAGGGAG TGGTTGTTTT CCTGCCTCAG CCCCACGCTC CTGCCGGTCC 24500 TGCACCTGCT GTAACCGTCG ATGTTGGTGC CAGGTGCCCA CCTGGGAAGG ATGCTGTGCA GGGGGCTTGC 24570 CAAACTTTGG TGGGTTTCAG AAGCCCCAGG CACTTGTGGC AGGCACAATT ACAGCCCCTC CCCAAAGATG 24640 CCCACGTCCT TCTCCTGGAA CCTGTGAATG TGTCACCCGC AAGGCAGAGG CTGGTGAAGG CTGCAGGTGG 24710 AATCACGGCT GCCAGTCAGC CGATCTTAAG GTCATCCTGG ATTATCTGGT GGGCCTGATA TGGCCACAAG 24780 GGTCCCTAGA AGTGAGAGAG GCAGGCAGGG GAGAGTCAGA GAGGGGACGT GAGAAGGACC ACTGGCCACT 24850 GCTGGCTTTG AGATGGAGGA GGAGGTCCCC AGCCAAGGAA TGGGGGCAGC CGCTCCATGC TGGAAAAGCA 24920 AGCAATCCTC CCCGGTCCTG AGGGCACACG GCCCTGCCCA CGCCTCGATT TCAGGCCAGT GGGACCTGTT 24990 TCAGCTTTCC GGCCTCCAGA GCTGTAAGAT GATGCGTTTG TGTTCAGCCA CTAAGCTGCA GTGATTCGTC 25060 ACAGCAGCAA ATGGAATAGC AGTACAGGGA AATGAATACA GGGACAGTTC TCAGAGTGAC TCTCAGCCCA 25130 CCCCTGGG 25138

Example 5

Comparison of the above-described genomic hTC sequence and the sequence of the hTC cDNA (FIG. 6; corresponding to SEQ ID NO 2) made it possible to elucidate the exon-intron structure of the hITC gene. The genomic organization of the hTC gene is illustrated diagrammatically in FIG. 7. The coding region of the hTC gene is composed of 16 exons which var′ in size between 6′ bp and 1354 bp (see Table 1). Exon 1 contains the translation start codon ATG. The translation stop codon TGA and the 3-untranslated region lie on exon 16 (FIG. 8). No possible polyadenylation signal (AATAAA) was found either in exon 16 or in the 3195 bp of the following 3′-flanking region. The exon-intron transitions were determined on the basis of the consensus sequence  5′-Exon         Intron                    3′-Exon Pre-mRNA A/C   A  G | G   T  A/G  A  ... N C   A  G | G Frequency (%) 70    60 80 100 100 95   70      80 100 100 60 and listed in Table 1. With the exception of the 5′ splice site between exon 15 and intron 15, all the exon-intron transitions are in accord with the published (Shapiro and Senapathy, 1987) splice consensus sequence. The sizes of the introns are between 104 bp and 8616 bp. Since only part of intron 6 was isolated, it is not possible to determine the precise length of the hTC gene. Based on the part sequence of ˜4660 bp, which was obtained from intron 6, the minimum size of the hTERT gene is 37 kb.

Introns 1-5 and the 5′ region of intron 6, are contained in contig 1:

Intron 1: bp 11493-11596 (SEQ ID NO 4);

Intron 2: bp 12951-21566 (SEQ ID NO 5);

Intron 3: bp 21763-23851 (SEQ ID NO 6);

Intron 4: bp 24033-24719 (SEQ ID NO 7);

Intron 5: bp 24900-25393 (SEQ ID NO 8);

5′ region of intron 6: bp 25550-26414 (SEQ ID NO 9).

The 3′ region of intron 6, and introns 7-15, are located in contig 2 at the following positions:

3′ region of intron 6: bp 1-3782 (SEQ ID NO 10);

Intron 7: bp 3879-4858 (SEQ ID NO 11),

Intron 8: bp 4945-7429 (SEQ ID NO 12);

Intron 9: bp 7544-9527 (SEQ ID NO 13);

Intron 10: bp 9600-11470 (SEQ ID NO 14);

Intron 11: bp 11660- 15460 (SEQ ID NO 15;

Intron 12: bp 15588-16467 (SEQ ID NO 16);

Intron 13: bp 16530-19715 (SEQ ID NO 17);

Intron 14: 19841-20621 (SEQ ID NO 18);

Intron 15: 20760-21295 (SEQ ID NO 19).

The 3′-untranscribed region is also located in contig 2 at position 21960-25138 (SEQ ID NO 20).

The individual sequences of the abovementioned introns are as follows: Intron 1 GTGGGCCTCCCCGGGGTCGGCGTCCGGCTGGGGTTGAGGGCGGCCGGGGGGAACCAGCGACATGCGGAGAGCAGCGCAGG (SEQ ID NO 4) CGACTCAGGGCGCTTCCCCCGCAG Intron 2 GTGAGGAGGTGGTGGCCGTCGAGGGCCCAGGCCCCAGAGCTGAATGCAGTAGGGGCTCAGAAAAGGGGGCAGGCAGAGCC (SEQ ID NO 5) CTGGTCCTCCTGTCTCCATCGTCACGTGGGCACACGTGGCTTTTCGCTCAGGACGTCGAGTGGACACGGTGATCTCTGCC TCTGCTCTCCCTCCTGTCCAGTTTGCATAAACTTACGAGGTTCACCTTCACGTTTTGATGGACACGCGGTTTCCAGGCGC CGAGGCCAGAGCAGTGAACAGAGGAGGCTGGGCGCGGCAGTGGAGCCGGGTTGCCGGCAATGGGGAGAAGTGTCTGGAAG CACAGACGCTCTGGCGAGGGTGCCTGCAGGTTACCTATAATCCTCTTCGCAATTTCAAGGGTGGGAATGAGAGGTGGGGA CGAGAACCCCCTCTTCCTGGGGGTGGGAGGTAAGGGTTTTGCAGGTGCACGTGGTCAGCCAATATGCAGGTTTGTGTTTA AGATTTAATTGTGTGTTGACGGCCAGGTGCGGTGGCTCACGCCGGTAATCCCAGCACTTTGGGAAGCTGAGGCAGGTGGA TCACCTGAGGTCAGGAGTTTGAGACCAGCCTGACCAACATGGTGAAACCCTATCTGTACTAAAAATACAAAAATTAGCTG GGCATGGTGGTGTGTGCCTGTAATCCCAGCTACTTGGGAGGCTGAGGCAGGAGAATCACTTGAACCCAGGAGGCGGAGGC TGCAGTGAGCTGAGATTGTGCCATTGTACTCCAGCCTGGGCGACAAGAGTGAAACTCTGTCTTTAAAAAAAAAAAGTGTT CGTTGATTGTGCCAGGACAGGGTAGAGGGAGGGAGATAAGACTGTTCTCCAGCACAGATCCTGGTCCCATCTTTAGGTAT GAAGAGGGCCACATGGGAGCAGAGGACAGCAGATGGCTCCACCTGCTGAGGAAGGGACAGTGTTTGTGGGTGTTCAGGGG ATGGTGCTGCTGGGCCCTGCCGTGTCCCCACCCTGTTTTTCTGGATTTGATGTTGAGGAACCTCCGCTCCAGCCCCCTTT TGGCTCCCAGTGCTCCCAGGCCCTACCGTGGCAGCTAGAAGAAGTCCCGATTTCACCCCCTCCCCACAAACTCCCAAGAC ATGTAAGACTTCCGGCCATGCAGACAAGGAGGGTGACCTTCTTGGGGCTCTTTTTTTTCTTTTTTTCTTTTTATGGTGGC AAAAGTCATATAACATGAGATTGGCACTCCTAACACCGTTTTCTGTGTACAGTGCAGAATTGCTAACTCGGCGGTGTTTA CAGCAGGTTGCTTGAAATGCTGCGTCTTGCGTGACTGGAAGTCCCTACCCATCGAACGGCAGCTGCCTCACACCTGCTGC GGCTCAGGTGGACCACGCCGAGTCAGATAAGCGTCATGCAACCCAGTTTTGCTTTTTGTGCTCCAGCTTCCTTCGTTGAG GAGAGTTTGAGTTCTCTGATCAGGACTCTGCCTGTCATTGCTGTTCTCTGACTTCAGATGAGGTCACAATCTGCCCCTGG CTTATGCAGGGAGTGAGGCGTGGTCCCCGGGTGTCCCTGTCACGTGCAGGGTGAGTGAGGCGTTGCCCCCAGGTGTCCCT GTCACGTGTAGGGTGAGTGAGGCGCGGCCCCCGGGTGTCCCTGTCCCGTGCAGCGTGATTGAGGTGTGGCCCCCGGGTGT CCCTGTCACGTGTAGGGTGAGTGAGGCGCCATCCCCGGGTGTCCCTGTCACGTGTAGGGTGAGTGAGGCGTGGTCCCCGG GTGTCCCTGTCCCGTGCAGGGTGAGTGAGGCACTGTCCCCGGGTGTCCCTGTCACGTGCAGGGTGAGTGAGGCGCGGTCC CCGGGTGTCCCTCTCAGGTGTAGGGTGAGTGAGGCGCGGCCCCAGGGTGTCCCTGTCACGTGTAGGGTGAGTGAGGCACC GTCCCTGGGTGTCCCTCCCAGGTATAGGGTGAGTGAGGCACTGTCCCCGGGTGTCCCTGTCACGTGCAGGGTGAGTGAGG CGCGGCCCCCGGGTGTCCCTCTCAGGTGCAGGGTGAGTGAGGCGCTGTCCCTGGGTGTCCCTGTCTCGTGTAGGGTGAGT GAGGCTCTGTCCCCAGGTGTCCTTGGCGTTTGCTCACTTGAGCTTGCTCCTGAATGTTTGCTCTTTCTATAGCCACAGCT GCGCCGGTTGCCCATTGCCTGGGTAGATGGTGCAGGCGCAGTGCTGGTCCCCAAGCCTATCTTTTCTGATGCTCGGCTCT TCTTGGTCACCTCTCCGTTCCATTTTGCTACGGGGACACGGGACTGCAGGCTCTCGCCTCCCGCGTGCGAGGCACTGCAG CCACAGCTTCAGGTCCGCTTGCCTCTGTTGGGCCTGGCTTGCTCACCACGTGCCCGCCACATGCATGCTGCCAATACTCC TCTCCCAGCTTGTCTCATGCCGAGGCTGGACTCTGGGCTGCCTGTGTCTGCTGCCACGTGTTGCTGGAGACATCCCAGAA AGGGTTCTCTGTGCCCTGAAGGAAAGCAAGTCACCCCAGCCCCCTCACTTGTCCTGTTTTCTCCCAAGCTGCCCCTCTGC TTGGCCCCCTTGGGTGGGTGGCAACGCTTGTCACCTTATTCTGGGCACCTGCCGCTCATTGCTTAGGCTGGGCTCTGCCT CCAGTCGCCCCCTCACATGGATTGACGTCCAGCCACAGGTTGGAGTGTCTCTGTCTGTCTCCTGCTCTGAGACCCACGTG GAGGGCCGGTGTCTCCGCCAGCCTTCGTCAGACTTCCCTCTTGGGTCTTAGTTTTGAATTTCACTGATTTACCTCTGACG TTTCTATCTCTCCATTGTATGCTTTTTCTTGGTTTATTCTTTCATTCCTTTTCTAGCTTCTTAGTTTAGTCATGCCTTTC CCTCTAAGTGCTGCCTTACCTGCACCCTGTGTTTTGATGTGAAGTAATCTCAACATCAGCCACTTTCAAGTGTTCTTAAA ATACTTCAAAGTGTTAATACTTCTTTTAAGTATTCTTATTCTGTGATTTTTTTCTTCGTGCACGCTGTGTTTTGACGTGA AATCATTTTGATATCAGTGACTTTTAAGTATTCTTTAGCTTATTCTGTGATTTCTTTGAGCAGTGAGTTATTTGAACACT GTTTATGTTCAAGATATGTAGAGTATCAAGATACGTAGAGTATTCTAAGTTATCATTCTATTATTGATTTCTAACTCAGT TGTGTAGTGGTCTGTATAATACCAATTATTTGAAGTTTGCGGAGCCTTGCTTTGTGATCTAGTGTGTGCATGGTTTCCAG AACTGTCCATTGTAAATTTGACATCCTGTCAATAGTGGGCATGCATGTTCACTATATCCAGCTTATTAAGGTCCAGTGCA AAGCTTCTGTCTCCTTCTAGATGCATGAAATTCCAAGAAGGAGGCCATAGTCCCTCACCTGGGGGATGGGTCTGTTCATT TCTTCTCGTTTGGTAGCATTTATGTGAGGCATTGTTAGGTGCATGCACGTGGTAGAATTTTTATCTTCCTGATGAGTGAA TCTTTTGGAGACTTCTATGTCTCTAGTAATCTAGTAATTCTTTTTTTAAATTGCTCTTAGTACTGCCACACTGGGCTTCT TTTGATTAGTATTTTCCTGCTGTGTCTGTTTTCTGCCTTTAATTTATATATATATATATATTTTTTTTTTTTTTGAGACA GAGTCTTGGTCTGTCGCCCAGGGTGAGTGCAGTGGTGTGATCACAGGTCAGTGTAACTTTTACCTTCTGGCCTGAGCCGT CCTCTCACCTCAGCCTCCTGAGTAGCTGGAACTGCAGACACGCACCGCTACACCTGGCTAATTTTTAAATTTTTTCTGGA GACAGGGTCTTGCTGTGTTGCCCAGGCTGGTCTCAAACTCTTGGACTCAAGGGATCCATCTACCTCGGCTTCCCAAAGTG CTGAATTACAGGCATGAGCCACCATGTCTGGCCTAATTTTCAACACTTTTATATTCTTATAGTGTGGGTATGTCCTGTTA ACAGCATGTAGGTGAATTTCCAATCCAGTCTGACAGTCGTTGTTTAACTGGATAACCTGATTTATTTTCATTTTTTTGTC ACTAGAGACCCGCCTGGTGCACTCTGATTCTCCACTTGCCTGTTGCATGTCCTCGTTCCCTTGTTTCTCACCACCTCTTG GGTTGCCATGTGCGTTTCCTGCCGAGTGTGTGTTGATCCTCTCGTTGCCTCCTGGTCACTGGGCATTTGCTTTTATTTCT CTTTGCTTAGTGTTACCCCCTGATCTTTTTATTGTCGTTGTTTGCTTTTGTTTATTGAGACAGTCTCACTCTGTCACCCA GGCTGGAGTGTAATGGCACAATCTCGGCTCACTGCAACCTCTGCCTCCTCGGTTCAAGCAGTTCTCATTCCTCAACCTCA TGAGTAGCTGGGATTACAGGCGCCCACCACCACGCCTGGCTAATTTTTGTATTTTTAGTAGAGATAGGCTTTCACCATGT TGGCCAGGCTGGTCTCAAACTCCTGACCTCAAGTGATCTGCCCGCCTTGGCCTCCCACAGTGCTGGGATTACAGGTGCAA GCCACCGTGCCCGGCATACCTTGATCTTTTAAAATGAAGTCTGAAACATTGCTACCCTTGTCCTGAGCAATAAGACCCTT AGTGTATTTTAGCTCTGGCCACCCCCCAGCCTGTGTGCTGTTTTCCCTGCTGACTTAGTTCTATCTCAGGCATCTTGACA CCCCCACAAGCTAAGCATTATTAATATTGTTTTCCGTGTTGAGTGTTTCTGTAGCTTTGCCCCCGCCCTGCTTTTCCTCC TTTGTTCCCCGTCTGTCTTCTGTCTCAGGCCCGCCGTCTGGGGTCCCCTTCCTTGTCCTTTGCGTGGTTCTTCTGTCTTG TTATTGCTGGTAAACCCCAGCTTTACCTGTGCTGGCCTCCATGGCATCTAGCGACGTCCGGGGACCTCTGCTTATGATGC ACAGATGAAGATGTGGAGACTCACGAGGAGGGCGGTCATCTTGGCCCGTGAGTGTCTGGAGCACCACGTGGCCAGCGTTC CTTAGCCAGTGAGTGACAGCAACGTCCGCTCGGCCTGGGTTCAGCCTGGAAAACCCCAGGCATGTCGGGGTCTGGTGGCT CCGCGGTGTCGAGTTTGAAATCGCGCAAACCTGCGGTGTGGCGCCAGCTCTGACGGTGCTGCCTGGCGGGGGAGTGTCTG CTTCCTCCCTTCTGCTTGGGAACCAGGACAAAGGATGAGGCTCCGAGCCGTTGTCGCCCAACAGGAGCATGACGTGAGCC ATGTGGATAATTTTAAAATTTCTAGGCTGGGCGCGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGCGGG TGGATCACGAGGTCAGGAGGTCGAGACCATCCTGGCCAACATGATGAAACCCCATCTGTACTAAAAACACAAAAATTAGC TGGGCGTGGTGGCGGGTGCCTGTAATCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATTGCTTGAACCTGGGAGTTGGAA GTTGCAGTGAGCCGACATTGCACCACTGCACTCCAGCCTGGCAACACAGCGAGACTCTGTCTCAAAAAAAAAAAAAAAAA AAAAAAAAAAAATTCTAGTAGCCACATTAAAAAAGTAAAAAAGAAAAGGTGAAATTAATGTAATAATAGATTTTACTGAA GCCCAGCATGTCCACACCTCATCATTTTAGGGTGTTATTGGTGGGAGCATCACTCACAGGACATTTGACATTTTTTGAGC TTTGTCTGCGGGATCCCGTGTGTAGGTCCCGTGCGTGGCCATCTCGGCCTGGACCTGCTGGGCTTCCCATGGCCATGGCT GTTGTACCAGATGGTGCAGGTCCGGGATGAGGTCGCCAGGCCCTCAGTGAGCTGGATGTGCAGTGTCCGGATGGTGCACG TCTGGGATGAGGTCGCCAGGCCCTGCTGTGAGCTGGATGTGTGGTGTCTGGATGGTGCAGGTCAGGGGTGAGGTCTCCAG GCCCTCGGTGAGCTGGAGGTATGGAGTCCGGATGATGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCTGTGAGCTGGATG TGTGGTGTCTGGATGGTGCAGGTCAGGGGTGAGGTCTCCAGGCCCTCGGTAAGCTGGAGGTATGGAGTCCGGATGATGCA GGTCCGGGGTGAGGTCGCCAGGCCCTGCTGTGAGCTGGATGTGTGGTGTCTGGATGGTGCAGGTCTGGGGTGAGGTCACC GGTCCGGGGTGAGGTCGCCAGGCCCTGCTGTGAGCTGGATGTGTGGTGTCTGGATGGTGCAGGTCTGGGGTGAGGTCACC AGGCCCTGCGGTGAGCTGGGTGTGCGGTGTCTGGATGGTGCAGGTCTGGAGTGAGGTCGCCAGACGGTGCCAGACCATGC GGTGAGCTGGATATGCGGTGTCCGGATGGTGCAGGTCTGGGGTGAGGTTGCCAGGCCCTGCTGTGAGTTGGATGTGGGGT GTCCGGATGCTGCAGGTCCGGTGTGAGGTCACCAGGCCCTGCTGTGAGCTGGATGTGTGGTGTCTGGATGGTGCAGGTCT GGGGTGAAGGTCGCCAGGCCCCTGCTTGTGAGCTGGATGTGTGGTGTCTGGATGGTGCAGGTCTGGAGTGAGGTCGCCAG GCCCTCGGTGAGCTGGATGTGCAGTGTCCAGATGGTGCAGGTCCGGGGTGAGGTCGCCAGACCCTGCGGTGAGCTGGATG TGCGGTGTCTGGATGGTGCAGGTCTGGAGTGAGGTCGCCAGGCCCTCGGTGAGCTGGATGTATGGAGTCCGGATGGTGCC GGTCCGGGGTGAGGTCGCCAGACCCTGCTGTGAGCTGGATGTGCGGTGTCTGGATGGTACAGGTCTGGAGTGAGGTCGCC AGACCCTGCTGTGAGCTGGATATGCGGTGTCCGGATGGTGCAGGTCAGGGGTGAGGTCTCCAGGCCCTCGGTGAGCTGGA GGTATGGAGTCCGGATGATGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCTGTGAACTGGATGTGCGGCGTCTGGATGGT GCAGGTCTGGGGTGTGGTCGCCAGGCCCTCGGTGAGCTGGAGGTATGGAGTCCGGATGATGCAGGTCCGGGGTGAGGTCG CCAGGCCCTGCTGTGAGCTGGATGTGCGGCGTCTGGATGGTGCAGGTCTGGGGTGTGGTCGCCAGGCCCTCGGTGAGCTG GAGGTATGGAGTCCGGATGATGCAGGTCCGGGGTGAGGTTGCCAGGCCCTGCTGTGAGCTGGATGTGCTGTATCCGGATG GTGCAGTCCGGGGTGAGGTCGCCAGGCCCTGCTGTGAGCTGGATGTGCTGTATCCGGATGGTGCAGGTCTGGGGTGAGGT CACCAGGCCCTGCGGTGAGCTGGTTGTGCGGTGTCCGGTTGCTGCAGGTCCGGGGTGAGTTCGCCAGGCCCTCGGTGAGC TGGATGTGCGGTGTCCCCGTGTCCGGATGGTGCAGGTCCAGGGTGAGGTCGCTAGGCCCTTGGTGGGCTGGATGTGCCGT GTCCGGATGGTGCAGGTCTGGGGTGAGGTCGCCAGGCCTTTGGTGAGCTGGATGTGCGGTGTCTGCATGGTGCAGGTCTG GGGTGAGGTCGCCAGGCCCTTGGTGGGCTGGATGTGTGGTGTCCGGATGGTGCAGGTCCGGCGTGAGGTCGCCAGGCCCT GCTGTGAGCTGGATGTGCGGTGTCTGGATGGTGCAGGTCCGGGGTGAGGTAGCCAAGGCCTTCGGTGAGCTGGATGTGGG GTGTCCGGATGGTGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCGGTTAGCTGGATATGCGGTGTCCGGATGGTGCAGGT CCGGGGTGAGGTCACCAGGCCCTGCGGTTAGCTGGATGTGCGGTGTCTGGATGGTGCAGGTCCGGGGTGAGGTCGCCAGG CCCTGCTGTGAGCTGGATGTGCTGTATCCGGATGGTGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCAGTGAGCTGGATG TGCTGTATCCGGATGGTGCAGGTCTGGCGTGAGGTCGCCAGGCCCTGCGGTTAGCTGGATATGCGGTGTCGGATGGTGCA GGTCCGGGGTGAGGTCACCAGGCCCTGCGGTTAGCTGGATGTGCGGTGTCCGGATGGTGCAGGTCTGGGGTGAGGTCGCC AGGCCCTGCTGTGAGCTGGATGTGCTGTATCCGGATGGTGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCGGTGAGCTGG ATGTGCTGTATCCGGATGGTGCAGGTCTGGCGTGAGGTCGCCAGGCCCTGCGGTGAGCTGGATGTGCAGTGTACGGATGG TGCAGGTCCGGGGTGAGGTCGCCAGGCCCTGCGGTGGGCTGTATGTGTGTTGTCTGGATGGTGCAGGTCCGGGGTGAGTT CGCCAGGCCCTGCGGTGAGCTGGATGTGTGGTGTCTGGATGCTGCAGGTCCGGGGTGAGTTCGCCAGGCCCTCGGTGAGC TGGATATGCGGTGTCCCCGTGTCCGAATGGTGCAGGTCCAGGGTGAGGTCGCCAGGCCCTTGGTGGGCTGGATGTGCCGT GTCCGGATGGTGCAGGTCTGGGGTGAGGTCGCCAGGCCCTTGGTGAGCTGGATGTGCGGTGTCCGGATGGTGCAGGTCCG GGGTGAGGTCACCAGGCCCTCGGTGATCTGGATGTGGCATGTCCTTCTCGTTTAAG Intron 3 GTACTGTATCCCCACCCCAGGCCTCTGCTTCTCGAAGTCCTGGAACACCAGCCCGGCCTCAGCATGCGCCTGTCTCCACT (SEQ ID NO 6) TGCCTGTGCTTCCCTGGCTGTGCAGCTCTGGGCTGGGAGCCAGGGGCCCCGTCACAGGCGTGGTCCAAGTGGATTCTGTG CAAGGCTCTGACTGCCTGGAGCTCACGTTCTCTTACTTGTAAAATCAGGAGTTTGTGCCAAGTGGTCTCTAGGGTTTGTA AAGCAGAAGGGATTTAAATTAGATGGAAACACTACCACTAGCCTCCTTGCCTTTCCCTGGGATGTGGGTCTGATTCTCTC TCTCTTTTTTTTTTCTTTTTTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGGAGTGCAGTGGCATAATCTTGGCTCACT GCAACCTCCACCTCCTGGGTTTAAGCGATTCACCAGCCTCAGCCTCCTAAGTAGCTGGGATTACAGGCACCTGCCACCAC GCCTGGCTAATTTTTGTACTTTTAGGAGAGACGGGGTTTCACCATGTTGGCCAGGCTGGTCTCGAACTCATGACCTCAGG TGATCCACCCACCTTCGCCTCCCAAAGTGCTGGGTTTACAGGCTAAGCCACCGTGCCCAGCCCCCGATTCTCTTTTAATT CATGCTGTTCTGTATGAATCTTCAATCTATTGGATTTAGGTCATGAGAGGATAAAATCCCACCCACTTGGCGACTCACTG CAGGGAGCACCTGTGCAGGGAGCACCTGGGGATAGGAGAGTTCCACCATGAGCTAACTTCTAGGTGGCTGCATTTGAATG GCTGTGAGATTTTGTCTGCAATGTTCGGCTGATGAGAGTGTGAGATTGTGACAGATTCAAGCTGGATTTGCATCAGTGAG GGACGGGAGCGCTGGTCTGGGAGATGCCAGCCTGGCTGAGCCCAGGCCATGGTATTAGCTTCTCCGTGTCCCGCCCAGGC TGACTGTGGAGGGCTTTAGTCAGAAGATCAGGGCTTCCCCAGCTCCCCTGCACACTCGAGTCCCTGGGGGGCCTTGTGAC ACCCCATGCCCCAAATCAGGATGTCTGCAGAGGGAGCTGGCAGCAGACCTCGTCAGAGGTAACACAGCCTCTGGGCTGGG GACCCCGACGTGGTGCTGGGGCCATTTCCTTGCATCTGGGGGAGGGTCAGGGCTTTCCCTGTGGGAACAAGTTAATACAC AATGCACCTTACTTAGACTTTACACGTATTTAATGGTGTGCGACCCAACATGGTCATTTGACCAGTATTTTGGAAAGAAT TTAATTGGGGTGACCGGAAGGAGCAGACAGACGTGGTGGTCCCCAAGATGCTCCTTGTCACTACTGGGACTGTTGTTCTG CCTGGGGGGCCTTGGAGGCCCCTCCTCCCTGGACAGGGTACCGTGCCTTTTCTACTCTGCTGGGCCTGCGGCCTGCGGTC AGGGCACCAGCTCCGGAGCACCCGCGGCCCCAGTGTCCACGGAGTGCCAGGCTGTCAGCCACAGATGCCCAGGTCCAGGT GTGGCCGCTCCAGCCCCCGTGCCCCCATGGGTGGTTTTGGGGGAAAAGGCCAAGGGCAGAGGTGTCAGGAGACTGGTGGG CTCATGAGAGCTGATTCTGCTCCTTGGCTGAGCTGCCCTGAGCAGCCTCTCCCGCCCTCTCCATCTGAAGGGATGTGGCT CTTTCTACCTGGGGGTCCTGCCTGGGGCCAGCCTTGGGCTACCCCAGTGGCTGTACCAGAGGGACAGGCATCCTGTGTGG AGGGGCATGGGTTCACGTGGCCCCAGATGCAGCCTGGGACCAGGCTCCCTGGTGCTGATGGTGGGACAGTCACCCTGGGG GTTGACCGCCGGACTGGGCGTCCCCAGGGTTGACTATAGGACCAGGTGTCCAGGTGCCCTGCAAGTAGAGGGGCTCTCAG AGGCGTCTGGCTGGCATGGGTGGACGTGGCCCCGGGCATGGCCTTCAGCGTGTGCTGCCGTGGGTGCCCTGAGCCCTCAC TGAGTCGGTGGGGGCTTGTGGCTTCCCGTGAGCTTCC

CCTAGTCTGTTGTCTGGCTGAGCAAGCGTCCTGAGGGGCTCT CTATTGCAG Intron 4 GTGGCTGTGCTTTGGTTTAACTTCCTTTTTAAACAGAAGTGCGTTTGAGCCCCACATTTGGTATCAGCTTAGATGAAGGG (SEQ ID NO 7) CCCGGAGGAGGGGCCACGGGACACAGCCAGGGCCATGGCACGGCGCCAACCCATTTGTGCGCACAGTGAGGTGGCCGAGG TGCCGGTGCCTCCAGAAAAGCAGCGTGGGGGTGTAGGGGGAGCTCCTGGGGCAGGGACAGGCTCTGAGGACCACAAGAAG CAGCCGGGCCAGGGCCTGGATGCAGCACGGCCCGAGGTCCTGGATCCGTGTCCTGCTGTGGTGCGCAGCCTCCGTGCGCT TCCGCTTACGGGGCCCGGGGACCAGGCCACGACTGCCAGGAGCCCACCGGGCTCTGAGGATCCTGGACCTTGCCCCACGG CTCCTGCACCCCACCCCTGTGGCTGCGGTGGCTGCGGTGACCCCGTCATCTGAGGAGAGTGTGGGGTGAGGTGGACAGAG GTGTGGCATGAGGATCCCGTGTGCAACACACATGCGGCCAGGAACCCGTTTCAAACAGGGTCTGAGGAAGCTGGGAGGGG TTCTAGGTCCCGGGTCTGGGTGGCTGGGGACACTGGGGAGGGGCTGCTTCTCCCCTGGGTCCCTATGGTGGGGTGGGCAC TTGGCCGGATCCACTTTCCTGACTGTCTCCCATGCTGTCCCCGCCAG Intron 5 GTGGGTGCCGGGGACCCCCGTGAGCAGCCCTGCTGGACCTTGGGAGTGGCTGCCTGATTGGCACCTCATGTTGGGTGGAG (SEQ ID NO 8) GAGGTACTCCTGGGTGGGCCGCAGGGAGTGCAGGTGACCCTGTCACTGTTGAGGACACACCTGGCACCTAGGGTGGAGGC CTTCAGCCTTTCCTGCAGCACATGGGGCCGACTGTGCACCCTGACTGCCCGGGCTCCTATTCCCAAGGAGGGTCCCACTG GATTCCAGTTTCCGTCAGAGAAGGAACCGCAACGGCTCAGCCACCAGGCCCCGGTGCCTTGCACCCCAGTCCTGAGCCAG GGGTCTCCTGTCCTGAGGCTCAGAGAGGGGACACAGCCCGCCCTGCCCTTGGGGTCTGGAGTGGTGGGGGTCAGAGAGAG AGTGGGGGACACCGCCAGGCCAGGCCCTGAGGGCAGAGGTGATGTCTGAGTTTCTGCGTGGCCACTGTCAGTCTCCTCGC CTCCACTCACACAG 5′-region intron 6 CTAAGGTTCACGTGTGATAGTCGTGTCCAGGATGTGTGTCTCTGGGATATGAATGTGTCTAGAATGCAGTCGTGTCTGTG (SEQ ID NO 9) ATGCGTTTCTGTCGTGGAGGTACTTCCATGATTTACACATCTGTGATATGCGTGTGTGGCACGTGTGTGTCGTGGTGCAT GTATCTGTGGCGTGCATATTTGTGGTGTGTGTGTGTGTGGCACGTGTGTGTCCATGGTGTGTGTGCCTGTGGTGTGCATG TGTGTGTGTCTGTGACACGTGCATGTTCATGCTGTGTGCTGCATGTCTGTGATGTGCCTATTTGTGGTGTGTGTGTGCAT GTGTCCGTGACATATGCGTGTCTATGGCATGGGTGTGTGTGGCCCCTTGGCCTTACTCCTTCCTCCTCCAGGCATGGTCC GCACCATTGTCCTCACGCTCTCGGGTGCTGGTTTGGGGAGCTCCACATTCAGGGTCCTCACTTCTAGCATGGGTGCCCCT GTCCTGTCACAGGGCTGGGCCTTGGAGACTGTAAGCCAGGTTTGAGAGGAGAGTAGGGATGCTGGTGGTACCTTCCTGGA CCCCTGGCACCCCCAGGACCCCAGTCTGGCCTATGCCGGCTCCATGAGATATAGGAAGGCTGATTCAGGCCTCGCTCCCC GGGACACACTCCTCCCAGAGCGGCCGGGGGCCTTGGGCCTCGGCAGGGGTGAAAGGGGCCCTGGGCTTGGGTTCCCACCC AGTGGTCATGAGCACGCTGGAGGGGTAAGCCCTCAAAGTCGTGCCAGGCCGGGGTGCAGAGGTGAAGAAGTATCCCTGGA GCCTTCGGTCTGGGGAGAGGCACATGTGGAAACCACAAGGACCTCTTTCTCTGACTTCTTGAGCT 3′-region intron 6 TGTGGGATTGGTTTTCATGTGTGGGATAGGTGGGGATCTGTGGGATTGGTTTTTATGAGTGGGGTAACACAGAGTTCAAG (SEQ ID NO 10) GCGAGCTTTCTTCCTGTAGTGGGTCTGCAGGTGCTCCAACAGCTTTATTGAGGAGACCATATCTTCCTTTGAACTATGGT CGGGTTTATAGTAAGTCAGGGGTGTGGAGGCCTCCCCTGGGCTCCCTGTTCTGTTTCTTCCACTCTGGGGTCGTGTGGTG CCTGCTGTGGTGTGTGGCCGGTGGGCAGGGCTTCCAGGCCTCCTTGTGTTCATTGGCCTGGATGTGGCCCTGGCTACGCT CCGTCCTTGGAATTCCCCTGCGAGTTGGAGGCTTTCTTTCTTTCTTTTTTTCTTTCTTTTTTTTTTTTTTTGATAACAGA GTCTCGCTCTTTTTTGCCCAGGCTGGAGTGGTTTGGCGTGATCTTGGCTCACTGCAACCTGTGCTTCCTGAGTTCAAGCA ATTCTCTTGCCTCAGCCTCCCAAGTAGCTGGAATTATAGGCGCCCACCACCATGCTGACTAATTTTTGCAATTTTAGTAG AGACGAGGTTTCTCCATGTTGGCCAGGCTGGTCTCGAACTCCTGACCTCAGGTGATCCTCCCACCTCGGCCTCCCAAAGT GCTGGGATGACAGGTGTGAACCGCCGCGCCCGGCCGAGACTCGCTTCCTGCAGCTTCCGTGAGATCTGCAGCGATAGCTG CCTGCAGCCTTGGTGCTGACAACCTCCGTTTTCCTTCTCCAGGTCTCGCTAGGGGTCTTTCCATTTCATGACTCTCTTCA CAGAAGAGTTTCACGTGTGCTGATTTCCCGGCTGTTTCCTGCGTAATTGGTGTCTGCTGTTTATCGATGGCCTCCTTCCA TTTCCTTTAGGCTTTGTTTATTGTTGTTTTTCCGGCTCCTTGAAGGAAAAGTTTCGATTATGGATGTTTGAACTTTCTTT TCTAAACAAGCATCTGAAGTTGCCGTTTTCCCTCTAAAGCAGGGATCCCGAGGCCCCTGGCTGTGGAGTGGCACCGGTCT GGGGCCTGTTAGGAACCCGGCGCACAGCGGGAGGCTAGGTGGGGTGTGGGGAGCCAGCGTTCCCGCCTGAGCCCCGCCCC TCTCAGATCAGCAGTGGCATGCGGTGCTCAGAGGCGCACACACCCTACTGAGAACTGTGCGTGAGAGGGGTCTAGATTCT GTGCTCCTTATGGGAATCTAATGCCTGATGATCTGAGGTGGAACCGTTTGCTCCCAAAACCATCCCCTTCCCCACTGCTG TCCTGTGGAAAAATCGTCTTCCACGAAACCAGTCCCTGGTACCACAATGGTTGGGGACCCTGTGCTAAAGACCTGCTTCA GCAGCCTCTCGTCAGTGTTGATATATTGGCTTTTCTGTGTTGAGTCCAGAATAATTACGGATTTCTGTGATGCTTTCCGC CGACCTCAGACCCATGGGCTATTTGTGGGCGTGTTGCCTGCTCCTGGGTTGGGAAGGGTGCAGGCCCCATGTACCTTCCT GTTACTGCCTTCCAGGTTGGTTCTCAGGGTTGAATCGTACTCGATGTGGTTTTAGCCCACGGCCCTGCCGCCAGCTCCTG GGGGCTGGGGAACATGCTGAAGCACAGAGTCACCGTGCGCGTCTTTTGATGCCTCACAAGCTCGAGGCCTCCTGTGTCCG TGTTAGTGTGTGTCACGTGCCTGCTCACATCCTGTCTTGGGGACGCAGGGGCTTAGCAGGTCCCGTAGTAAATGACAAGC GTCCTGGGGGAGTCTGCAGAATAGGAGGTGGGGGTGCCGGTCTCTCTCCCGCGTCTTCAGACTCTTCTCCTGCCTGTGCT GTGGCTGCACCTGCATCCCTGCAATCCCTCCAGCACTGGGCTGGAGAGGCCCGGGAGCTCGAGTGCCACTTGTGCCACGT GACTGTGGATGGCAGTCGGTCACGGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTTGGTCACAGGGGTCTGATGTGTG GTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGG ATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCGGTCGTG GGGTCTGATGTGGTGACTGTGGATGGCAGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATG TGGTGACTGTGGATGGCAGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACT GTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGG CGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGTGATCGGTCA CAGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGTGATCGGTCACAG GGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTTGGTCCCGGGGG TCTGATGTGTGGTGACTGTGGATGGCGATCGGTCACAGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCT GATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGGT GACTGTGGATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGAT GGCGGTTGGTCCCGGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGATGGCAG TCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGG TCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGT GGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGTGATCGGTCACAGGGGTCTGATGTGTGGT GACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGGTGACTGTGGAT GGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTAGGGTCTGATGTGTGGTGACTGTGGATGGCAGTCG GTCACAGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGG GGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGATGTGTGGTGACTGTGGATGGCGGTCGTGGGGTCTGAT GTGGTGACTGTGGATGGTGATCGGTCACAGGGGTCTGATGTGTGGTAGCTGCAGGTGGAGTCCCAGGTGTGTCTGTAGCT ACTTTGCGTCCTCGGCCCCCCGGCCCCCGTTTCCCAAACAGAAGCTTCCCAGGCGCTCTCTGGGCTTCATCCCGCCATCG GGCTTGGCCGCAGGTCCACACGTCCTGATCGGAAGAAACAAGTGCCCAGCTCTGGCCGGGGCAGGCCACATTTGTGGCTC ATGCCCTCTCCTCTGCCGGCAG Intron 7 GTCTGGGCACTGCCCTGCAGGGTTGGGCACGGACTCCCAGCAGTGGGTCCTCCCCTGGGCAATCACTGGGCTCATGACCG (SEQ ID NO 11) GACAGACTGTTGGCCCTGGGGGGCAGTGGGGGGAATGAGCTGTGATGGGGGCATGATGAGCTGTGTGCCTTGGCGAAATC TGAGCTGGGCCATGCCAGGCTGCGACAGCTGCTGCATTCAGGGACCTGCTCACGTTTGACTGCGCGGCCTCTCTCCAGTT CCGCAGTGCCTTTGTTCATGATTTGCTAAATGTCTTCTCTGCCAGTTTTGATCTTGAGGCCAAAGGAAAGGTGTCCCCCT CCTTTAGGAGGGCAGGCCATGTTTGAGCCGTGTCCTGCCCAGCTGGCCCCTCAGTGCTGGGTCTGAGGCCAAAGGAAACG TGTCCCCCTTCTTAGGAGGACGGGCCGTGTTTGAGCCACGCCCCGCTGAGCGGGCCTCTCAGTGCTGGGTCTGTCCACGT GGCCCTGTGGCCCTTTGCAGATGTGGTCTGTCCACGTGGCCCTGTGGCTCTTTGCAGATGCCTGTTAGCACTTGCTCGGC TCTAGGGGACAGTCGTGTCCACCGCATGAGGCTCAGAGACCTCTGGGCGAATTTCCTTGGCTCCCAGGGTGGGGGTGGAG GTGGCCTGGGCTGCTGGGACCCAGACCCTGTGCCCGGCAGCTGGGCAGCAACTCCTGGATCACATATGCCATCCGGGCCA CGGTGGGCTGTGTGGGTGTGAGCCCAGCTGGACCCACAGGTGGCCCAGAGGAGACGTTCTGTGTCACACACTCTGCCTAA GCCCATGTGTGTCTGCAGAGACTCGGCCCGGCCAGCCCACGATGGCCCTGCATTCCAGCCCAGCCCCGCACTTCATCACA AACACTGACCCCAAAAGGGACGGAGGGTCTTGGCCACGTGGTCCTGCCTGTCTCAGCACCCACCGGCTCACTCCCATGTG TCTCCCGTCTGCTTTCGCAG Intron 8 GTGAGTCAGGTGGCCAGGTGCCATTGCCCTGCGGGTGGCTGGGCGGGCTGGCAGGGCTTCTGCTCACCTCTCTCCTGCCC (SEQ ID NO 12) CTTCCCCACTGNCCTTCTGCCCGGGGCCACCAGAGTCTCCTTTTCTGGCCCCCGCCCCCTCCGGCTCCTGGGCTGCAGGC TCCCGAGGCCCCGGAAACATGGCTCGGCTTGCGGCAGCCGGAGCGGAGCAGGTGCCACACGAGGCCTGGAAATGGCAAGC GGGGTGTGGAGTTGCTCCTGCGTGGAGGACGAGGGGCGGGGGGTGTGTCTGGGTCAGGTGTGCGCCGAGCGTTTGAGCCT GCAGCTTGTCAGCTCCAAGTTACTACTGACGCTGGACACCCGGCTCTCACACGCTTGTATCTCTCTCTCCCGATACAAAA GGATTTTATCCGATTCTCATTCCTGTCCCTGTCGTGTGACCCCCGCGAGGGCGCGGGCTCTTCTCTCTGTGACTAGATTT CCCATCTGGAAAGTGCGGGGTTGACCGTGTAGTTTGCTCCTCTCGGGGGGCCTGTGGTGGCCATGGGGCAGGCGGCCTGG GAGAGCTGCCGTCACACAGCCACTGGGTGAGCCACACTCACGGTGGTAGAGCCACAGTGCCTGGTGCCACATCACGTCCT CTGGATTTTAAGTAAAACCACACACCTCCCGGCAGGCATCTGCCTGCGACCCTGTGTGTGCCTGGGGAGAGTGGTAGCAC GGAGGAAATTCGTGCACACTCAAGGTCATCAGCAAGGTCATCCGCAGTCAGGTGGAACGTGGAGGCCTCTCTCTGGGATC GTCTCCAGCGGATAAAGGACTGTGCACAGCTTCGGAAGCTTTTATTTAAAAATATAACTATTAATTATTGCATTATAAGT AATCACTAATGGTATCAGCAATTATAATATTTATTAAAGTATAATTAGAAATATTAAGTAGTACACACGTTCTGGAAAAA CACAAATTGCACATGGCAGCAGAGTGAATTTTGGCCGAGGGACACGTGTGCACATGTGTGTAAGCGGCCCCCAGGCCCAC AGAATTCGCTGACAAAGTCACCTCCCCAGAGAAGCCACCACGGGCCTCCTTCGTGGTCGTGAATTTTATTAAGATGGATC AAGTCACGTACCGTCCACGTGTGGCAGGGCTTTGGGGAATGTGAGGTGATGACTGCGTCCTCATGCCCTGACAGACAGGA GGTGACTGTGTCTGTCCTGTCCCTAGGACACGGACAGGCCCGAAGCTCTAGTCCCCATCGTGGTCCAGTTTGGCCTCTGA ATAAAAACGTCTTCAAAACCTGTTGCCCCAAAAACTAAGAACAGAGAGAGTTTCCCATCCCATGTGCTCACAGGGGCGTA TCTGCTTGCGTTGACTCGCTGGGCTGGCCGGACTCCTAGAGTTGGTGCGTGTGCTTCTGTGCAAAAAGTGCAGTCCTCTT GCCCATCACTGTGATATCTGCACCAGCAAGGAAAGCCTCTTTTCTTTTCTTTCTTTTTTTTTTTTTGAGACGGAACGTCA CTGTTGTCTGCCTGGGCTTGAGTGCAGTGGCGCGATCTCAACTCACTGCAACCTCCGCCTCCCGGGTTCCAGCATTTCTC CTGCCTCAGCCTCCCGAGCAGCTGAGATTACAGGCACCCACCCCCTGCGCCTGGCTAATTTTTGTATTTTTAGTAGAGAG GGGTTTTTGCCATGTTGGCCAGGCTGGTCTCGAACTCCTGACCTCAGGTGATCCACCCACCTCGGCCTCCCAAAGTGCTG GGATTACAGGTGTGAGCCATCACGCCCAGCCGGAAAGCCTCTTTTTAAGGTGACCACCTATAGCGCTTCCCGAAAATAAC AGGTCTTGTTTTTGCAGTAGGCTGCAAGCGTCTCTTAGCAACAGGAGTGGCGTCCTGTGGGCTCTGGGGATGGCTGAGGG TCGCGTGGCAGCCATGCCTTCTGTGTGCACCTTTAGGTTCCACGGGGCTATTCTGCTCTCACTGTTTGTCTGAAAACGCA CCCTTGGCATCCTTGTTTGGAGAGTTTCTGCTTCTCGTTGGTCATGCTGAAACTAGGGGCAAGGTTGTATCCGTTGGCGC GCAGCGGCTACATGTAGGGTCATGAGTCTTTCACCGTGGACAAATTCCTTGAAAAAAAAAAAAGGAGTCCGGTTAAGCAT TCATTCCGGGTCAAGTGTCTGGTTCTGTGAATAAACTCTAAGATTTAAGAAACCTTAATGAAAGAAAACCTTGATGATTC AGAGCAAGGATGTGGTCACACCTGTGGCTGGATCTGTTTCAGCCGCCCCAGTGCATGGTGAGAGTGGGGAGCAGGGATTG TTTGTTCAGAGGTCTCATCTGGTATGTTTCTGAGGTGTTTGCCGGCTGAATGGTAGACGTGTCGTTTGTGTGTATGAGGT TCTGTGTCTGTGTGTGGCTCGGTTTGAGTGTACGCATGTCCAGCACATGCCCTGCCCGTCTCTCACCTGTGTCTTCCCGC TCCAG Intron 9 CCGAGGCCTCCTCTTCCCCAGGGGGGCTTGGGTGGGGGTTGATTTGCTTTTGATGCATTCAGTGTTAATATTCCTGGTGC (SEQ ID NO 13) CCTGGAGACCATGACTGCTCTGTCTTGAGGAACCAGACAAGGTTGCAGCCCCTTCTTGGTACGAAGCCGCACGGGAGGGG TTGCACAGCCTGAGGACTGCGGGCTCCACGCAGGCTCTGTCCAGCGGCCATGTCCAGAGGCCTCAGGGCTCAGCAGGCGG GAGGGCCGCTGCCCTGCATGATGAGCATGTGAATTCAACACCGAGGAAGCACACCAGCTTCTGTCACGTCACCCAGGTTC CGTTAGGGTCCTTGGGGAGATGGGGCTGGTGCAGCCTGAGGCCCCACATCTCCCAGCAGGCCCTCGACAGGTGGCCTGGA CTGGGCGCCTCTTCAGCCCATTGCCCATCCCACTTGCATGGGGTCTACACCCAAGGACGCACACACCTAAATATCGTGCC AACCTAATGTGGTTCAACTCAGCTGGCTTTTATTGACAGCAGTTACTTTTTTTTTTTTAATACTTTAAGTTCTAGGGTAC ATGTGCACGACGTGCAGGTTAGTTACATATGTATACATGTGCCATGTTGGTGTGCTGCACCCATTAACTCATCATTTACA TTAGGTATATCTCCTAATGCTATCCCTCCCCACTCCCCCCATCCCATGACAGGCCCTGGTGTGTGATGTTCCCCACCCTG TGTCCAAGTGTTCTCATTGTTCAGTTCCCACCTGTGAGTGAGAACATGTGGTGTTTGGTTTTCTTTCCTTGCAATAGTTT GCTCAGAGTGATGGTTTCCAGCTTCGTCCATGTCCCTACAAAGGACATGAACTCATCCTTTTTTATGACTGCATAGTATT CCGTGGTGTATATGTGCCACATTTTCTTAATCCAGTCTATCATCGATGGACATTTGGGTTGGTTGCAAGTCTTTGCTACT GTGAATAGTGCCGCAATAAACATACGTGTGCATGTGTCTTTATAGCAGCATGATTTATAATCCTTTGGGTATATACCCAG TAATGGGATGGCTGGGTCAAATGGTATTTCTAGTTCTAGATCCTTGAGGAATCACCACACTGTCTTCCACAATGGTTGAA CTAGTTTACACTCCCACCAACAGTGTAAAAGTGTTCTGGTGCTGGAGAGGATGTGGACAGCAGTTATTTTTTTATGAAAA TAGTATCACTGAACAAGCAGACAGTTAGTGAAGGATGCGTCAGGAAGCCTGCAGGCCACACAGCCATTTCTCTCGAAGAC TCCGGGTTTTTCCTGTGCATCTTTTGAAACTCTAGCTCCAATTATAGCATGTACAGTGGATCAAGGTTCTTCTTCATTAA GGTTCAAGTTCTAGATTGAAATAAGTTTATGTAACAGAAACAAAAATTTCTTGTACACACAACTTGCTCTGGGATTTGGA GGAAAGTGTCCTCGAGCTGGCGGCACACTGGTCAGCCCTCTGGGACAGGATACCTCTGGCCCATGGTCATGGGGCGCTGG GCTTGGGCCTGAGGGTCACACAGTGCACCATGCCCAGCTTCCTGTGGATAGGATCTGGGTCTCGGATCATGCTGAGGACC ACAGCTGCCATGCTGGTAAAGGGCACCACGTGGCTCAGAGGGGGCGAGGTTCCCAGCCCCAGCTTTCTTACCGTCTTCAG GCTGATGGTAAACACTGAGTACTTATAATGAATGAGGAATTGCTGTAGCAGTTAACTGTAGAGAGCTCGTCTGTTGGAAA GAAATTTAAGTTTTTCATTTAACCGCTTTGGAGAATGTTACTTTATTTATGGCTGTGTAAATTGTTTGACATTCAGTCCC TCGTAGACAGATACTACGTAAAAAGTGTAAAGTTAACCTTGCTGTGTATTTTCCCTTATTTTAG Intron 10 GTGAGGCCCGTGCCGTGTGTCTGTGGGGACCTCCACAGCCTGTGGGCTTTGCAGTTGAGCCCCCCGTGTCCTGCCCCTGG (SEQ ID NO 14) CACCGCAGCGTTGTCTCTGCCAAGTCCTCTCTCTCTGCCGGTGCTGGATCCGCAAGAGCAGAGGCGCTTGGCCGTGCACC CAGGCCTGGGGGCGCAGGGGCACCTTCGGGAGGGAGTGGGTACCGTGCAGGCCCTGGTCCTGCAGAGACGCACCCAGGTT ACACACGTGGTGAGTGCAGGCGGTGACCTGGCTCCTGCTGCTCTTTGGAAAGTCAAGAGTGGCGGCTCCTGGGGCCCCAG TGAGACCCCCAGGAGCTGTGCACAGGGCCTGCAGGGCCGAGGCGGCAGCCTCCTCCCCAGGGTGCACCTGAGCCTGCGGA GAGCAGGAGCTGCTGAGTGAGCTGGCCCACAGCGTTCGCTGCGGTCACGTTCCTGCGTGGGGTTGTTTGGGATCGGTGGG AGAATTTGGATTTCCTGAGTGCTGCTGTCTTGAACCACGGAGATGGCTAGGAGTGGGTTTCAGAGTTGATTTTTGTGAAT CAAACTAAAATCAGGCACAGGGGACCTGGCCTCAGCACAGGGGATTGTCCCCCTGTGGTCCCCCTCAAGGGCGCCCACAG AGCCGGTGGGCTTGTTTTAAAGTGCGATTTGACGAGGGACGAGAAACCTTGAAAGCTGTAAAGGGAACCCTCAGAAAATG TGGCCGCCAGGGGTGGTTTCAGGTGCTTTGCTGGGCTGTGTTTGTGAAAACCCATTTGGACCCGCCCTCCAAGTCCACCC TCCAGGTCCACCCTCCAGGGCCGCCCTGGGCTGGGGGTATGCCTGGCGTTCCTTGTGCCGCAGCCCGGAGCACAGCAGGC TGTGCACATTTAAATCCACTAAGATTCACTCGGGGGGAGCCCAGGTCCCAAGCAACTGAGGGCTCAGGAGTCCTGAGGCT GCTGAGGGGACAGAGCAGACGGGGAACGCTGCTTCTGTGTGGCAAGTTCCTGAGGGTGCTGGCCAGGGAGGTGGCTCAGA GTGTATGTTGGGGTCCCACCGGGGGCAGAACTCTGTCTCTGATGAGTCGGCAGCCATGTAACAGGAAGGGGTGGCCACAG GGAGCTGGGAATGCACCAGGGGAGCTGCGCAGCTGGCCGAGGTCCCAGGGCCAGGCCACAGGAAGGGCAGGGGGACGCCC GGGGCCACAGCAGAGGCCGCAGGAAGGGAAGGGGATGCCCAGGCCAGAGCAGAGGCTACCGGGCACAGGGGGGCTCCCTG AGCTGGGTGAGCGAGGCTCATGACTCGGCGAGGGAACCTCCTTGACGTGAAGCTGACGACTGGTGTTGCCCAGCTCACAG CCCAGCCAGGTCCCGCGCCTGAGCAGGAACTCAGAACCCTCCCCTTTGTCTAAAGCACAGCAGATGCCTTCAGGGCATCT AGGAGAAAACAGGCAAAGTCGTTGAGAAACGTCTTAAAAGAAGGTGGGATGGTGGCAATTTCTTGTCCAGATCTTAGTCT GCCCCGGACCACAGATGAGTCTATAACGGGATTGTGGTGTTGCCATGGGGACACATGAGATGGACCATCACACAGGCCAC TGGGGCTGCACCTCCCATCTGAGTCCTGGCTGTCCCGGGTCCAGGCCAGGTTCTTGCATGCTCACCTACCTC

GCTGCCC GGGAGACAGGGAAAGCACCCCGAAGTCTGGAGCAGGGCTGGGTCCAGGCTCCTCAGAGCTCCTGCCAGGCCCAGCACCCT GCTCCAAATCACCACTTCTCTGGGGTTTTCCAAAGCATTTAACAAGGGTGTCAGGTTACCTCCTGGGTGACGGCCCCGCA TCCTGGGGCTGACATTGCCCCTCTGCCTTAG Intron 11 GTGAGCGCACCTGGCCGGAAGTGGAGCCTGTGCCCGGCTGGGGCAGGTGCTGCTGCAGGGCCGTTGCGTCCACCTCTGCT (SEQ ID NO 15) TCCGTGTGGGGCAGGCGACTGCCAATCCCAAAGGGTCAGAGGCCACAGGGTGCCCCTCGTCCCATCTGGGGCTGAGCAGA AATGCATCTTTCTGTGGGAGTGAGGGTGCTCACAACGGGAGCAGTTTTCTGTGCTATTTTGGTAAAAGGAAATGGTGCAC CAGACCTGGGTGCACTGAGGTGTCTTCAGAAAGCAGTCTGGATCCGAACCCAAGACGCCCGGGCCCTGCTGGGCGTGAGT CTCTCAAACCCGAACACAGGGGCCCTGCTGGGCATGAGTCCCTCTGAACCCGAGACCCTGGGGCCCTGCTGGGCGTGAGT CTCTCCGAACCCAGAGACTTCAGGGCCCTTTTGGGCGTGAGTCTCTCCGCTGTGAGCCCCACACTCCAAGGCTCATCCAC AGTCTACAGGATGCCATGAGTTCATGATCACGTGTGACCCATCAGGGGACAGGGCCATGGTGTGGGGGGGGTCTCTACAA AATTCTGGGGTCTTGTTTCCCCAGAGCCCGAGAGCTCAAGGCCCCGTCTCAGGCTCAGACACAAATGAATTGAAGATGGA CACAGATGCAGAAATCTGTGCTGTTTCTTTTATGAATAAAAAGTATCAACATTCCAGGCAGGGCAAGGTGGCTCACACCT ATAATCCCAGCACTTTGGGAGGCCGAGGTGGGTGGATCACTTGAGGCCAGGAGTTTGAGGCCAACCTAACCAACATAGTG AAATTCCATTTCTACTTAAAAAATACAAAAATTAGCCTGGCCTGGTGGCACACGCCTGTAGTCCCCGCTATGCGGGAGGC TGAGGCAGGAGAATCATTTGAACCCAGGAGGCAGAGGTTGCAGTGAGCCGAGATCACACCACTGCACTCCAGCCTGGGCA ACAGAGTGAGACTTCATCTTAAAAAAAAAAAAAAAAGTATCAGCATTCCAAAACCATAGTGGACAGGTGTTTTTTTATTC TGTCCTTCGATAATATTTACTGGTGCTGTGCTAGAGGCCGGAACTGGGGGTGCCTTCCTCTGAAAGGCACACCTTCATGG GAAGAGAAATAAGTGGTGAATGGTTGTTAAACCAGAGGTTTAAACTGGGGTCCTGTCGTTCTGAGTTAACAGTCCAGATC TGGACTTTGCCTCTTTCCAGAATGCTCCCTGGGGTTTGCTTCATGGGGGAGCAGCAGGTGTGGACACCCTCGTGATGGGG GAGCAGCAGGTGCAGACGCCCTCATGATGGGGGAGTGGCAGGTGCAGACACCCTTGTGCATGGTGCCCAGCATGTCCCTG TTGCAGCTCCCTCCCCACAAGGATGCCGGTCTCCTGTGCTCCCCACAGTCCCTGCTTCCCTCTCACAGCCTTACCTGGTC CTGGCCTCCACTGGCTTTGTCTGCATGATTTCCACATTTCCTGGGCTCCCAGCACCTCTTCGCCTCTCCCAGGCACCTCT GCAGTCCTGGCCATACCAGTCAGCTGTGAACTGTCCACTGCTTATTTTGCTCCCCATGAAATGTATTTTTTAGGACAGGC ACCCCTGGTTCCAGCCTCTGGCACAGCATCAGTGAATGTTATTGAAGGACAAAGGACAGACAAACAAATCAGGAAAATGG GTTCTCTCTAAACACATTGCAAAGCCACAGAGGCTAGTGCAGGATGGGTGGGCATCAGGTCATCAGATGTGGGTCCAATG CCAGAATATTCTGTGCTCCCAAAGGCCACTTGGTCAGAGTGTGTGCTTGCAGAGGTGGCTCTAAAAGCTCAGCAGTGGAG GCAGTGGTTCGCCATACTCAGGGTGAACTCACATCCTCTGTGTCTGAAGTATACAGCAGAGGCTTGAAGGGCATCTGGGA GAAGAAAACAGGCAAAATGATTAAGAAAAGTGAAAAAGGAAAAGTGGTAAGATGGGAATTTTCTTGTCCAGATTTTAGTC TCCCAAACCACAGCTCAGATGGTAGAATGTGGTCAGAACTGATGGACAGAACAATAGAACAAAACGGAAGCCCTATCTCT CAGAAACGTGTGTTAATGTGGTATGTGGCACAGCTGATGGAAAAGAGAGTGTGTGTGTAATTTTTTTTTCTGAGAAAACT GACTGGAAGCAAATAAGTTGTGTCTTTACAGCATATACCAGAGCAGATTCTAGGTAGAAGAGGAGACACATGCAAACAAC ACCAGCAACAGAAATAAAACAAAAGACTCAAAGGGAAGGGAGGTGAACGTTCCCTGGTTTGGTGTTGGGGAAGGACACAC AGGGAGGCGGATGAAACCACTGAGGCAACCGGCATTGCTTTCACTGCAGAGAAACTCAGCTTGCCTGAGCCACAGTGAAA ATGGCCATTCCCTGGAGCGTTTGTGCACGTGATTTATTTAAGGCGCCCTGTGAGGTCCTGCACATTCATCCTCTCACTTT GTTCTCCTAACCACCTGAGAGGTAGAGGAGGAAAGGCTCCAGGGGAGCAGCCGCCCTTGGTCACCCAGCTGGCAAAGGGC ATGCATGATTGCAGCCTGGCCTCCTGCTCCGGGGCCCTTGCTCTGCCCGAGGACCCCACACAAGTCAGACCCATAGGCTC AGGGTGAGCCGGAGCCCAAGGTCGTGTTGGGGATGGCTGTGAAAGAAGAAATGGACGTCTGATGCACACTTGGGAAGGTC CTACCAGCAGCGTCAAAGAAATGCATGTGAAACTGACAGCGAGACCCATCCCTCAAAGAAACGCACGTGAAACTGATGGC GAGACCTGTCCCCATCCCTCATGCTGGCTCCTTTTCTGGGCTTGCCAAGAGCCAGCATCAGGTTGAGGCAAGCTGGAAAG ACTTTTCTGGAAAGCAGCTTGTTTGCATGGAAGTCCTCACAATGTCCTGTGTCTTCCCAGTAATTCCACTTCTGAAGTGA CCAGACATTATCACGGGTCTTATTTACCATTTCCAGTGTTCCAGGCAGGGGGACTTGCCACAGCAAGTCACGAACCTGCC CAAATACAGGGCTAAGGAGATATTATGCATCACAAAACTTGCTCTGCCATTAAACATTTTTCAAAGAATTTTTGAAGAAT GTTTAATGGCACAAAACGTTTATTTCAATGTAGCAGTGTTCAAAGCTGGCTGTAAAAGAACACACCCCAGGAGCCTGCCG TGAATGTCATGTGTGTTCATCTTTGGACATGGACATACATGGGCAGTGAGTGGTGGTGAGGCCCTGGAGGACATCGGTGG GATGCCTCCATCCTGCCCCTCTGGAGACACCATGTGTGCCACGTGCACTCACTGGAGCCCTGTTTAGCTGGTGCCACCTG GCTCTTCCATCCCTGAGATTCAAACACAGTGAGATTCCCCACGCCCAACTCAGTGTTCTCCCACAAAAAACCTGAGTCAC ACCTGTGTTCACTCGAGGGACGCCCGGGAGCCAGGGCTCCACAGTTTATTATGTGTTTTTGGCTGAGTTATGTGCAGATC TCATCAGGGCAGATGATGAGTGCACAAACACGGCCGTGCGAGGTTTGGATACACTCAACATCACTAGCCAGGTCCTGGTG GAGTTTGGTCATGCAGAGTCTGGATGGCATGTAGCATTTGGAGTCCATGGAGTGAGCACCCAGCCCCCTCGGGCTGCAGC GCATGCCCCAGGCAGGACAAGGAAGCGGGAGGAAGGCAGGAGGCTCTTTGGAGCAAGCTTTGCAGGAGGGGGCTGGGTGT GGGGCAGGCACCTGTGTCTGACATTCCCCCCTGTGTCTCAG Intron 12 GTGAGCAGGCTGATGGTCAGCACAGAGTTCAGAGTTCAGGAGGTGTGTGCGCAAGTATGTGTGTGTGTGTGTGCGCGCGT (SEQ ID NO 16) GCCTGCAAGGCTGATGGTGACTGGCTGCACGTAAGAGTGCACATGTACGCATATACACGTGAGCACATACATGTGTGCAT GTGTGTACATGAAGGCATGGCAGTGTGTGCACAGGTGTGCAAGGGCACAAGTGTGTGCACATGCGAATGCACACCTGACA TGCATGTGTGTTCGTGCACAGTCGTGTGGGCATTCACGTGAGGTGCATGCGTGTGGGTGTGCAGTGTGAGTAGCATGTGT GCACATAACATGTATTGAGGGGTCCTCGTGTTCACCCCGCTAGGTCCTCAGCACCAGTGCCACTCCTTAGAGGATGAGAC GGGGTCCCACGCCTTGGTGGGCTGAGGCTCTGAAGCTGCAGCCCTGAGGGCATTGTCCCATCTGGGCATCCGCGTCCACT CCCTCTCCTGTGGGCTTCTGTGTCCACTCCCCCTCTCCTGTGGGCATTTACATCCACTCCACTCCCTCTCTCCTGTGGGC ATCCGCGTCCACTCCCCCTCTCTGTGGGCATCTGCGTCCACCTCCCCTCTCTGTGGGCATTTGCGTCCACTCCCTCTCCT GGTTCCTTCCTGTCTTGGCCGAGCCTCGGGGGCAGGCAGATGACACAGAGTCTTGACTCGCCCAGGGTGGTTCGCAGCTG CCGGGTGAGGGCCAGGCCGGATTTCACTGGGAAGAGGGATAGTTTCTTGTCAAAATGTTCCTCTTTCTTGTTCCATCTGA ATGGATGATAAAGCAAAAAGTAAAAACTTAAAATCCCAGAGAGGTTTCTACCGTTTCTCACTCTTTCTTGGCGACTCTAG Intron 13 GTGAGCCGCCACCAAGGGGTGCAGGCCCAGCCTCCAGGGACCCTCCGCGCTCTGCTCACCTCTGACCCGGGGCTTCACCT (SEQ ID NO 17) TGGAACTCCTGGGTTTTAGGGGCAAGGAATGTCTTACGTTTTCAGTGGTGCTGCTGCCTGTGCACAGTTCTGTTCGCGTG GCTCTGTGCAAAGCACCTGTTCTCCATCTCTGGGTAGTGGTAGGAGCCGGTGTGGCCCCAGGTGTCCCCACTGTGCCTGT GCACTGGCCGTGGGACGTCATGGAGGCCATCCCAGGGCAGCAGGGGCATGGGGTAAAGAGATGTTTATGGGGAGTCTTAG CAGAGGAGGCTGGGAAGGTGTCTGAACAGTAGATGGGAGATCAGATGCCCGGAGGATTTGGGGTCTCAGCAAAGAGGGCC GAGGTGGGTGCAGGTGAGGGTCGCTGGCCCCACCCCCGGGAAGGTGCAGCAGAGCTGTGGCTCCCCACACAGCCCGGCCA GCACCTGTGCTCTGGGCATGGCTGTGCTCCTGGAACGTTCCCTGTCCTGGCTGGTCAGGGGGTGCCCCTGCCAAGAATCG ACAACTTTATCACAGAGGGAAGGGCCAATCTGTGGAGGCCACAGGGCCAGCTTCTGCCTGGAGTCAGGGCAGGTGGTGGC ACAAGCCTCGGGGCTGTACCAAAGGGCAGTCGGGCACCACAGGCCCGGGCCTCCACCTCAACAGGCCTCCCGAGCCACTG GGAGCTGAATGCCAGGAGGCCGAAGCCCTCGCCCCATGAGGGCTGAGAAGGACTGTGAGCATTTGTGTTACCCAGGGCCG AGAGCCACAGCTGCATGTTACCGCCTTTGCACCAGCTCCAGAGGCTTGGGACCAGGCTGTCTCAGTTCCAGGGTGCGTCC GGCTCAGACCGCCCTCCTCTCTGCCTTCTCTCTCTGCCTCAAATCTTCCCTCGTTTGCATCTCCCTGACGCGTGCCTGGG CCCTCGTGCAAGCTGCTTGACTCCTTTCCGGAAACCCTTGGGGTGTGCTGGATACAGGTGCCACTGAGGACTGGAGGTGT CTGACACTGTGGTTGACCCCAGGGTCCAGCTGGCGTGCTTGGGGCCTCCTTGGGCCATGATGAGGTCAGAGGAGTTTTCC CAGGTGAAAACTCCTGGGAAACTCCCAGGGCCATGTGACCTGCCACCTGCTCCTCCCATATTCAGCTCAGTCTTGTCCTC ATTTCCCCACCAGGGTCTCTAGCTCCGAGGAGCTCCCGTAGAGGGCCTGGGCTCAGGGCAGGGCGGCTGAGTTTCCCCAC CCATGTGGGGACCCTTGGGTAGTCGCTTGATTGGGTAGCCCTGAGGAGGCCGAGATGCGATGGGCCACGGGCCGTTTCCA AACACAGAGTCAGGCACGTGGAAGGCCCAGGAATCCCCTTCCCTCGAGGCAGGAGTGGGAGAACGGAGAGCTGGGCCCCG ATTTCACGGCAGCCAGGCTGCAGTGGGCGAGGCTGTGGTGGTCCACGTGGCGCTGGGGGCGGGGTCTGATTCAAATCCGC TGGGGCTCGGCCTTCCTGGCCCGTGCTGGCCGCGCCTCCACACGGGCTTGGGGTGGACGCCCCGACCTCTAGCAGGTGGC TATTTCTCCCTTTGGAAGAGAGCCCCTCACCCATGCTAGGTGTTTCCCTCCTGGGTCAGGAGCGTGGCCGTGTGGCAACC CCGGGACCTTAGGCTTATTTATTTGTTTAAAAACATTCTGGGCCTGGCTTCCGTTGTTGCTAAATGGGGAAAAGACATCC CACCTCAGCAGAGTTACTGAGAGGCTGAAACCGGGGTGCTGGCTTGACTGGTGTGATCTCAGGTCATTCCAGAAGTGGCT CAGGAAGTCAGTGAGACCAGGTACATGGGGGGCTCAGGCAGTGGGTGAGATGAGGTACACGGGGGGCTCAGGCAGTGGGT GAGGCCAGGTACATGGGGGGCTCAGGCACTGGGTGAGATGAGGTACACGGGGGGCTCAGGCAGAGGGTCAGACCAGGTAC ACGGGGGCTCTGATCACACGCACATATGAGCACATGTGCACATGTGCTGTTTCATGGTAGCCAGGTCTGTGCACACCTGC CCCAAAGTCCCAGGAAGCTGAGAGGCCAAAGATGGAGGCTGACAGGGCTGGCGCGGTGGCTCACACCTGTAGTCCCAGCA CTTTGGGAGGCCGAGGCGAGAGGATCCCTTGAGCCCAGGAGTTTAAGACCAGCCTGAGCAACATAGTAGAACCCCATCTC TATGAAAAATAAAAACAAAAATTAGCTGAACATGGTGGTGTGCGCCTGTAGTTCCAATACTTGGGAGGCTGAAGTGGGAG GATCACTTGAGCCCAGGAGGTGGAAGCTGCAGTGAGCTGAGATTGCACCACTGTACTGCAGCCTGGGTGACAGAGTGAGA GCCCATCTCAACAACAACAAAGAAGACTGACAAATGCAGTTTCTTGGAAAGAAACATTTAGTAGGAACTTAACCTACACA CAGAAGCCAAGTCGGTGTCTCGGTGTCAGTGAGATGAGATGATGGGTCCTCACACCATCACCCCAGACCCAGGGTTTATG CACCACAGGGGCGGGTGGCTCAGAAGGGATGCGCAGGACGTTGATATACGATGACATCAAGGTTGTCTGACGAAGGGCAG GATTCATGATAAGTACCTGCTGGTACACAAGGAACAATGGATAAACTGGAAACCTTAGAGGCCTTCCCGGAACAGGGGCT AATCAGAAGCCAGCATGGGGGGCTGGCATCCAGGATGGAGCTGCTTCAGCCTCCACATGCGTGTTCATACAGATGGTGCA CAGAAACGCAGTGTACCTGTGCACACACAGACACGCAGCTACTCGCACACACAAGCACACACACAGACATGCATGCATGC ATCCGTGTGTGTGCACCTGTGCCCATGAGGAAACCCATGCATGTGCATTCATGCACGCACACAGGCACCGGTGGGCCCAT GCCCACACCCACGAGCACCGTCTGATTAGGAGGCCTTTCCTCTGACGCTGTCCGCCATCCTCTCAG Intron 14 GTATGTGCCGGTGCCTGGCCTCAGTGGCAGCAGTGCCTGCCTGCTGGTGTTAGTGTGTCAGGAGACTGAGTGAATCTGGG (WEQ ID NO 18) CTTAGGAAGTTCTTACCCCTTTTCGCATCAGGAAGTGGTTTAACCCAACCACTGTCAGGCTCGTCTGCCCGCCCTCTCGT GGGGTGAGCAGAGCACCTGATGGAAGGGACAGGAGCTGTCTGGGAGCTGCCATCCTTCCCACCTTGCTCTGCCTGGGGAA GCGCTGGGGGGCCTGGTCTCTCCTGTTTGCCCCATGGTGGGATTTGGGGGGCCTGGCCTCTCCTGTTTGCCCTGTGGTGG GATTGGGCTGTCTCCCGTCCATGGCACTTAGGGCCCTTGTGCAAACCCAGGCCAAGGGCTTAGGAGGAGGCCAGGCCCAG GCTACCCCACCCCTCTCAGGAGCAGAGGCCGCGTATCACCACGACAGAGCCCCGCGCCGTCCTCTGCTTCCCAGTCACCG TCCTCTGCCCCTGGACACTTTGTCCAGCATCAGGGAGGTTTCTGATCCGTCTGAAATTCAAGCCATGTCGAACCTGCGGT CCTGAGCTTAACAGCTTCTACTTTCTGTTCTTTCTGTGTTGTGGAAATTTCACCTGGAGAAGCCGAAGAAAACATTTCTG TCGTGACTCCTGCGGTGCTTGGGTCGGGACAGCCAGAGATGGAGCCACCCCGCAGACCGTCGGGTGTGGGCAGCTTTCCG GTGTCTCCTGGGAGGGGAGCTGGGCTGGGCCTGTGACTCCTCAGCCTCTGTTTTCCCCCAG Intron 15 GCAAGTGTGGGTGGAGGCCAGTGCGGGCCCCACCTGCCCAGGGGTCATCCTTGAACGCCCTGTGTGGGGCGAGCAGCCTC (SEQ ID NO 19) AGATGCTGCTGAAGTGCAGACGCCCCCGGGCCTGACCCTGGGGGCCTGGAGCCACGCTGGCAGCCCTATGTGATTAAACG CTGGTGTCCCCAGGCCACGGAGCCTGGCAGGGTCCCCAACTTCTTGAACCCCTGCTTCCCATCTCAGGGGCGATGGCTCC CCACGCTTGGGAGCCTTCTGACCCCTGACCTGTGTCCTCTCACAGCCTCTTCCCTGGCTGCTGCCCTGAGCTCCTGGGGT CCTGAGCAAGTTCTCTCCCCGCCCCGCCGCTCCAGCGTCACTGGGCTGCCTGTCTGCTCGCCCCGGTGGAGGGGTGTCTG TCCCTTCACTGAGGTTCCCACCAGCCAGGGCCACGAGGTGCAGGCCCTGCCTGCCCGGCCACCCACACGTCCTAGGAGGG TTGGAGGATGCCACCTCTCGCCTCTTCTGGAACGGAGTCTGATTTTGGCCCCGCAG 3′-untranscribed region ATCTCATGTTTGAATCCTAATGTGCACTGCATAGACACCACTGTATGCAATTACAGAAGCCTGTGAGTGAACGGGGTGGT (SEQ ID NO 20) GGTCAGTGCGGGCCCATGGCCTGGCTGTGCATTTACGGAAGTCTATGAGTGAATGGGGTTGTGGTCAGTGCGGGCCCATG GCCTGGCTGGGCCTGGGAGGTTTCTGATGCTGTGAGGCAGGAGGGGAAGGAGGGTAGGGGATAGACAGTGGGAGCCCCCA CCCTGGAAGACATAACAGTAAGTCCAGGCCCGAAGGGCAGCAGGGATGCTGGGGGCCCAGCTTGGGCGGCGGGGATGATG GAGGGCCTGGCCAGGGTGGCAGGGATGATGGGGGCCCCAGCTGGGGTGGCAGGGGTGATGGGGGGGGCTGGTCTGGGTGG CGGGGAAGATGGGGAAGCCTGGCTGGGCCCCCTCCTCCCCTGCCTCCCACCTGCAGCCGTGGATCCGGATGTGCTTCCCT GGTGCACATCCTCTGGGCCATCAGCTTTCATGGAGGTGGGGGGCAGGGGCATGACACCATCCTGTATAAAATCCAGGATT CCTCCTCCTGAACGCCCCAACTCAGGTTGAAAGTCACATTCCGCCTCTGGCCATTCTCTTAAGAGTAGACCAGGATTCTG ATCTCTGAAGGGTGGGTAGGGTGGGGCAGTGGAGGGTGTGGACACAGGAGGCTTCAGGGTGGGGCTGGTGATGCTCTCTC ATCCTCTTATCATCTCCCAGTCTCATCTCTCATCCTCTTATCATCTCCCAGTCTCATCTGTCTTCCTCTTATCTCCCAGT CTCATCTGTCATCCTCTTACCATCTCCCAGTCTCATCTCTTATCCTCTTATCTCCTAGTCTCATCCAGACTTACCTCCCA GGGCGGGTGCCAGGCTCGCAGTGGAGCTGGACATACGTCCTTCCTCAGGCAGAAGGAACTGGAAGGATTGCAGAGAACAG GAGGGGCGGCTCAGAGGGACGCAGTCTTGGGGTGAAGAAACAGCCCCTCCTCAGAAGTTGCCTTGGGCCACACGAAACCG AGGGCCCTGCGTGAGTGGCTCCAGAGCCTTCCAGCAGGTCCCTGGTGGGGCCTTATGGTATGGCCGGGTCCTACTGAGTG CACCTTGGACAGGGCTTCTGGTTTGAGTGCAGCCCGGACGTGCCTGGTGTCGGGGTGGGGGCTTATGGCCACTGGATATG GCGTCATTTATTGCTGCTGCTTCAGAGAATGTCTGAGTGACCGAGCCTAATGTGTATGGTGGGCCCAAGTCCACAGACTG TGTCGTAAATGCACTCTGGTGCCTGGAGCCCCCGTATAGGAGCTGTGAGGAAGGAGGGGCTCTTGGCAGCCGGCCTGGGG GCGCCTTTGCCCTGCAAACTGGAAGGGAGCGGCCCCGGGCGCCGTGGGCGGACGACCTCAAGTGAGAGGTTGGACAGAAC AGGGCGGGGACTTCCCAGGAGCAGAGGCCGCTGCTCAGGCACACCTGGGTTTGAATCACAGACCAACaGGTCAGGCCATT GTTCAGCTATCCATCTTCTACAAAGCTCCAGATTCCTGTTTCTCCGGGTGTTTTTTGTTGAAATTTTACTCAGGATTACT TATATTTTTTGCTAAAGTATTAGACCCTTAAAAAAGGTATTTGCTTTGATATGGCTTAACTCACTAAGCACCTACTTTAT TTGTCTGTTTTTATTTATTATTATTATTATTATTAGAGATGGTGTCTACTCTGTCACCCAGGTTGTTAGTGCAGTGGCAC AGTCATGGCTCGCTGTAGCCGCAAACCCCCAGGCTCAAGTGATCCTCCGGCCTCAGCTTCCCAGAGTGCTGGGATTACAG GTGTGAGCCACTGCCCTTGCCTGGCACTTTTAAAAACCACTATGTAAGGTCAGGTCCAGTGGCTTCCACACCTGTCATCC CAGTAGTTTGGGAAGCCGAGGCAGAAGGATTGTCTGAGGCCAGGAGTTTGAGACCAGCATGGGTAACATAGGGAGACCCC ATCTCTACAAAAAATGCAAAAAGTTATCCGGGCGTGGGGTCCAGCATCTGTAGTCCCAGCTGCTCGGGAGGCTGAGTGGG AGGATCGCTTGAGCCCGGGAGGTCATGGCTGCAGTGAGCTGTGATTGTACCATCGCACTCCAGCCTGGGCAACAGAGTGA GACCCTGTCTCAAAAAAAAAAAAAAAAAAAGAAGGAGAAGGAGAAGAGAAGAAGAAGGAAGAAGGAAAGAGAAGAAGAAG GAAGAAGGAAGAAAGAAGGAGAAGGAGGCCTGCTAGGTGCTAGGTAGACTGTCAAATCTCAGAGCAAAATGAAAATAACA AAGTTTTAAAGGGAAAGAAAAACCCCAGCTCTTTGGACTTCCTTAGGCCTGAACTTCATCTCAAGCAGCTTCCTTCCACA GACAAGCGTGTATGGAGCGAGTGAGTTCAAAGCAGAAAGGGAGGAGAAGCAGGCAAGGGTGGAGGCTGTGGGTGACACCA GCCAGGACCCCTGAAAGGGAGTGGTTGTTTTCCTGCCTCAGCCCCACGCTCCTGCCGGTCCTGCACCTGCTGTAACCGTC GATGTTGGTGCCAGGTGCCCACCTGGGAAGGATGCTGTGCAGGGGGCTTGCCAAACTTTGGTGGGTTTCAGAAGCCCCAG GCACTTGTGGCAGGCACAATTACAGCCCCTCCCCAAAGATGCCCACGTCCTTCTCCTGGAACCTGTGAATGTGTCACCCG CAAGGCAGAGGCTGGTGAAGGCTGCAGGTGGAATCACGGCTGCCAGTCAGCCGATCTTAAGGTCATCCTGGATTATCTGG TGGGCCTGATATGGCCACAAGGGTCCCTAGAAGTGAGAGAGGGAGGCAGGGGAGAGTCAGAGAGGGGACGTGAGAAGGAC CACTGGCCACTGCTGGCTTTGAGATGGAGGAGGGGGTCCCCAGCCAAGGAATGGGGGCAGCCGCTCCATGCTGGAAAAGC AAGCAATCCTCCCCGGTCCTGAGGGCACACGGCCCTGCCCACGCCTCGATTTCAGGCCAGTGGGACCTGTTTCAGCTTTC CGGCCTCCAGAGCTGTAAGATGATGCGTTTGTGTTCAGCCACTAAGCTGCAGTGATTCGTCACAGCAGCAAATGGAATAG CAGTACAGGGAAATGAATACAGGGACAGTTCTCAGAGTGACTCTCAGCCCACCCCTGGG

Characterization of the exons showed, interestingly, that the functionally important hTC protein domains which are described in our Patent Application PCT/EP/98/03469 are arranged on separate exons. The telomerase-characteristic T motif is located on exon 3. The RT (reverse transcriptase) motifs 1-7, which are important for the catalytic function of the telomerase, are located on the following exons: RT motifs 1 and 2 on exon 4, RT motif 4 on exon 9, RT motif 5 on exon 10, and RT motifs 6 and 7 on exon 11. RT motif 3 is shared by exons 5 and 6 (see FIG. 8).

Elucidation of the exon-intron structure of the hTC gene also shows that the four deletions or insertion variants of the hTC CDNA which were described in our Patent Application PCT/EP/98/03469, as well as three additional hTC insertion variants which are described in the literature (Kilian et al., 1997), in all probability represent alternative splicing products. As shown in FIG. 8, the splicing variants can be divided into two groups: deletion variants and insertion variants.

The hTC variants in the deletion group lack specific sequence segments. The 36 bp in-frame deletion in variant DEL1 in all probability results from using an alternative 3′ splice acceptor sequence in exon 6, resulting in a part of RT motif 3 being lost. In variant DEL2, the normal 5′ splice donor and 3′ splice acceptor sequences of introns 6, 7 and 8 are not used. Instead exon 6 is fused directly to exon 9, resulting in a displacement arising in the open reading frame and a stop codon appearing in exon 10. Variant Del3 is a combination of variants 1 and 2.

The insertion variant group is characterized by the insertion of intron sequences which lead to premature cessation of translation. Instead of the 5′ splice donor sequence of intron 5, which is normally used, use is made, in variant INS1, of an alternative, 3′-located splice site, resulting in the insertion of the first 38 bp from intron 4 between exon 4 and exon 5. The insertion, in variant INS2, of a region of the 11 sequence likewise results from using an alternative 5′ splice donor sequence in intron 11. since this variant was only described inadequately in the literature (Kilian et al., 1997), it is not possible to determine the precise alternative 5′ splice donor sequence in this variant. The insertion of intron 14 sequences between exon 14 and exon 15 in variant INS3 comes from using an alternative 3′ splice acceptor sequence, resulting in the 3′ part of intron 14 not being spliced.

The hTC variant INS4 (variante 4), which is described in our Patent Application PCT/EP/98/03469, is characterized by exon 15, and the 5′ part region of exon 16, being replaced by the first 600 bp of intron 14. This variant can be attributed to the use of an alternative internal 5′ splice donor sequence in intron 14 and an alternative 3′ splice acceptor sequence in exon 16, resulting in an altered C terminus.

The in vivo generation of hTC protein variants which are probably non-functional and which could interfere with the function of the complete hTC protein constitutes a possible mechanism, in addition to transcription regulation, for controlling hTC protein function. The function of the hTC splicing variants is not yet known. Although most of these variants presumably encode proteins without reverse transcriptase activity, they could nevertheless play a crucial role as transdominant-negative telomerase regulators by, for example, competing for interaction with important binding partners.

The search for possible transcription factor binding sites was carried out using the ,,find pattern” algorithm from the Genetics Computer Group (Madison, USA) GCG Sequence Analysis program package. This resulted in the identification of a variety of potential binding sites for transcription factors in the nucleotide sequence of intron 2, which binding sites are listed in Tab. 2. In addition, an SpI binding site was found in intron 1 (pos. 43), and a c-Myc binding site was found in the 5′-untranslated region (cDNA position 29-34, cf. FIG. 6).

Example 6

In order to ascertain the start point(s) of hTC transcription in HL 60 cells, the 5′ end of the hTC mRNA was determined by means of primer extension analysis.

2 μg of polyA⁺ RNA from HL-60 cells were denaturated at 65° C. for 10 min. 1 μl of RNasin (3040 U/ml) and 0.3-1 pmol of radioactively labelled primer (5′GTTAAGTTGTAGCTTACACTGGTTCTC 3′; 2.5-8×10⁵ cpm) were added for primer annealing, and the whole was incubated, at 37° C. for 30 min, in a total volume of 20 μl. After the addition of 10 μl of 5xreverse transcriptase buffer (from Gibco-BRL), 2 μl of 10 mM dNTPs, 2 μl RNasin (see above), 5 μl of 0.1 M DTT (from Gibco-BRL) 2 μl of ThermoScript RT (15 U/μl; from Gibco-BRL) and 9 μl of DEPC-treated water, primer extension took place, at 58° C. for 1 h, in a total volume [lacuna]. The reaction was stopped by adding 4 μl of 0.5 M EDTA, pH 8.0, and the RNA was degraded, at 37° C. for 30 min, after having added 1 μl of RNaseA (10 mg/ml). 2.5 μg of sheared calf thymus DNA and′ 100 μl of TE were then added, and the mixture was extracted once with 150 μl of phenol/chloroform (1:1). The DNA was precipitated, at −70° C. for 45 min, after adding 15 μl of 3 M Na acetate and 450 μl of ethanol, and then centrifuged at 14,000 rpm for 15 min. The precipitate was washed once with 70% ethanol, dried in air and dissolved in 8 μl of sequencing stop solution. After 5 min of denaturation at 80° C., the samples were loaded onto a 6% polyacrylamide gel and fractionated electrophoretically (Ausubel et al., 1987) (FIG. 5).

In this connection, a main transcription start site was identified which is located 1767 bp 5′ of the ATG start codon of the hTC cDNA sequence (nucleotide position 3346 in FIG. 4). In addition to this, the nucleotide sequence around this main transcription start (TTA⁻¹TTGT) represents an initiator element (Inr), which, in 6 out of 7 nucleotides, matches the consensus motif (PyPyA₊₁Na/tPyPy) (Smale, 1997) of an initiator element.

It was not possible to identify any unambiguous TATA box in the immediate vicinity of the experimentally identified main transcription start, which means that the hTC promoter has probably to be classified in the family of TATA-less promoters (Smale, 1997). However, a potential TATA box from nucleotide position 1306 to nucleotide position 1311 (FIG. 4) was found by means of bioinformatics analysis. The subsidiary transcription starts which were additionally observed around the main transcription start have also been described in the case of other TATA-less promoters (Geng and Johnson, 1993), for example in the strongly regulated promoters of some cell cycle genes (Wick et al., 1995).

Example 7

In addition to the start point of the hTC transcript which was described in Example 6 and identified in HL60 cells, a further transcription start region was also identified in HL60 cells. With the aid of RT-PCR analyses, the region of the hTC gene transcription start in HL60 cells was localized to bp -60 to bp -105.

The cDNA for this was synthesized using a First Strand cDNA Synthesis kit (Clontech), in accordance with the manufacturer's instructions, and employing 0.4 μg of HL60 cell polyA RNA (Clontech) and the gene-specific primer GSP13 (5′-CCTCCAAAGAGGTGGCTTCTTCGGC-3′, cDNA position 920-897). In a final volume of 50 μl, 10 pmol dNTP mix were added to 1 μl of cDNA, and a PCR reaction was carried out in 1×PCR reaction buffer F (PCR-Optimizer kit from InVitrogen) and using one unit of platinum Taq DNA polymerase (from Gibco/BRL). 10 pmol of each of the 5′ and 3′ primers defined below were added as primers. The PCR was carried out in 3 steps. A two-minute denaturation at 94° C. was followed by 36 PCR cycles in which the DNA was first of all denatured at 94° C. for 45 sec and, after that, the primers were annealed, and the DNA chain was extended at 68° C. for 5 min. The cycles were concluded by a chain extension at 68° C. for 10 min. In all, six different PCR primers (primer HTRT5B: 5′-CGCAGCCACTACCGCGAGGTGC-3′ cDNA position 105 to 126; primer C5S: 5′-CTGCGTCCTGCTGCGCACGTGGGAAGC-3′, 5′-flanking region -49 to -23; primer PRO-TEST1: 5′-CTCGCGGCGCGAGTTTCAGGCAG-3′, 5′-flanking region -74 to -52; primer PRO-TEST2: 5′-CCAGCCCCTCCCCTTCCTTTCC-3′, 5′-flanking region -112 to -91; primer PRO-TEST4: 5′-CCAGCTCCGCCTCCTCCGCGC-3′, 5′-flanking region -191 to -171; primer RP-3A: 5′-CTAGGCCGATTCGACCTCTCTCC-3′, 5′-flanking region -427 to -405) were combined with the 3′ PCR primer C5Rback (5′-GTCCCAGGGCACGCACACCAG-3′, cDNA position 245 to 225). Genomic DNA was also employed for the PCR, as a control, in addition to the Oligo dT- and GSP13-primed cDNAs. As FIG. 9 shows, a PCR product was only obtained with the primer combinations HTRT5B-C5Rback, C5S-C5Rback and PRO-TEST1-C5Rback, indicating that the start point for hTC transcription lies in the region between bp-60 and bp-105.

Example 8

Several extremely GC-rich regions, so-called CpG Islands, are located in the isolated 5′-flanking region, of about 11.2 kb in size, of the hTC gene. One CpG Island, having a GC content of >70%, extends from bp -1214 into intron 2. Two further GC-rich regions having a GC content of >60% extend from bp -3872 to bp -3113 and from bp -5363 to bp -3941, respectively. The positions of the CpG Islands are shown graphically in FIG. 11.

The search for possible transcription factor binding sites was carried out using the “Find Pattern” algorithm from the Genetics Computer Group (Madison, USA) GCG Sequence Analysis program package. This resulted in the identification of a variety of potential binding sites in the region up to -900 bp upstream of the translation start codon ATG: five Sp1 binding sites, one c-Myc binding site, and one CCAC box (FIG. 10). In addition, a CCAAT box and a second c-Myc binding site were found at positions -1788 and -3995, respectively, of the 5′-flanking region.

Example 9

In order to analyse the activity of the hTC promoter, PCR amplification was used to generate four hTC promoter sequence segments of differing length, which segments were cloned into the Promega vector pGL2 5′ in front of the luciferase reporter gene. The 8.5 kb SacI fragment which was subcloned from phage clone P12 was selected as the DNA source for the PCR amplification. In a final volume of 50 μl, 10 pmol of dNTP mix were added to 35 ng of this DNA, and a PCR reaction was carried out in 1×PCR reaction buffer (PCR-Optimizer kit from InVitrogen) and using one unit of platinum Taq DNA polymerase (from Gibco/BRL). In each case 20 pmol of the 5′ and 3′ primers which are defined below were added as primers. The PCR was carried out in three steps. A two-minute denaturation at 94° C. was followed by 30 PCR cycles in which the DNA was first of all denaturated at 94° C. for 45 sec, after which the primers were annealed, and the DNA chain was extended, at 68° C. for 5 min. The cycles were concluded by a chain extension at 68° C. for 10 min. The selected 3′ PCR primer was in each case the primer PK-3A (5′-GCAAGCTTGACGCAGCGCTGCCTGAAACTCG-3′, position 43 to -65), which primer recognizes a sequence region 42 bp upstream of the ATG START codon. A promoter fragment of 4051 bp in size (NPK8) was amplified by combining the PK-3A primers with the 5′ PCR primer PK-5B (5′-CCAGATCTCTGGAACACAGAGTGGCAGTTTCC-3′, position 4093 to -4070). Combining the pair of primers PK-3A and PK-5C (5′-CCAGATCTGCATGAAGTGTGTGGGGATTTGCAG-3′, position -3120 to -3096) led to the amplification of a promoter fragment of 3078 bp in size (NPK15). Use of the primer combination PK-3A and PK-5D (5′-GGAGATCTGATCTTGGCTTACTGCAGCCTCTG-3′, position -2110 to -2087) amplified a promoter fragment of 2068 bp in size (NPK22). Finally, using the primer combination PK-3A and PK-5E (5′-GGAGATCTGTCTGGATTCCTGGGAAGTCCTCA-3′, position - 1125 to -1102) led to the amplification of a promoter fragment of 1083 bp in size (NPK27).

The PK-3A primer contains a HindIII recognition sequence. The different 5′ primers contain a BglII recognition sequence.

The resulting PCR products were purified using the Qiagen QIA quick spin PCR purification kit, in accordance with the manufacturer's instructions, and then digested with the restriction enzymes BglII and HindIII. The pGL2 promoter vector was digested with the same restriction enzymes, and the SV40 promoter contained in this vector was released and removed. The PCR promoter fragments ligated into the vector, which was then transformed into competent DH5α bacteria (from Gibco/BRL). DNA for the promoter activity analyses, which are described below, was isolated from transformed bacterial clones using the Qiagen plasmid kit.

Example 10

The activity of the hTC promoter was analysed in transient transfections in eukaryotic cells.

All the work with eukaryotic cells was carried out at a sterile workstation. CHO-K1 and HEK 293 cells were obtained from the American Type Culture collection.

CHO-K1 cells were kept in DMEM Nut Mix F-12 cell culture medium (from Gibco-BRL, order number: 21331-020) containing 0.15% streptomycin/penicillin, 2 mM glutamine and 10% FCS (from Gibco-BR1).

HEK 293 cells were cultured in DMOD cell culture medium (from Gibco-BRL, order number: 41965-039) containing 0.15% streptomycin/penicillin, 2 mM glutamine and 10% FCS (from Gibco-BRL).

CHO-K1 and HEK 293 cells were cultured at 37° C. in a water-saturated atmosphere while being gassed with 5% CO₂. When the cell lawn was confluent, the medium was sucked off, after which the cells were washed with PBS (100 mM KH₂PO₄ pH 7.2; 150 mM NaCl) and released by adding a trypsin-EDTA solution (from Gibco-BRL). The trypsin was inactivated by adding medium and the cell count was determined using a Neubauer counting chamber in order to plate out the cells at the desired density.

For the transfection, in each case 2×10⁵ HEK 293 cells were plated out, per well, in a 24-well cell culture plate. The HEK 293 medium was removed after 3 hours. For the transfection, up to 2.5 μg of plasmid DNA, 1 μg of a CMV β-Gal plasmid construct (from Stratagene, order number: 200388), 200 μl of serum-free medium and 10 μl of transfection reagent (DOTAP from Boehringer Mannheim) were incubated at room temperature for 15 minutes and then dropped uniformly onto the HEK 293 cells. 1.5 ml of medium were added after 3 hours. Tbf medium was changed after 20 hours. After a further 24 hours, the cells were harvested for determining the luciferase activity and the β-Gal activity. For this, the cells were lysed, at room temperature for 15 minutes, in the cell culture lysis reagent (25 mM Tris [nH 7.8] containing H₃PO₄; 2 mM CDTA; 2 mM DTT; 10% glycerol; 1% Triton X-100). Twenty μl of this cell lysate were mixed with 100 μl of luciferase assay buffer (20 mM Tricin; 1.07 mM (MgCO₃)₄ Mg(OH)₂.5H₂O; 2.67 mM MgSO₄; 0.1 mM EDTA; 33.3 mM DTT; 270 μM coenzyme A; 470 μM luciferin, 530 μM ATP), and the light generated by the luciferase was measured.

In order to measure the β-galactosidase activity, equal quantities of cell lysate and β-galactosidase assay buffer (100 mM sodium phosphate buffer, pH 7.3; 1 mM MgCl₂; 50 mM β-mercaptoethanol; 0.665 mg of ONPG/ml) were incubated at 37° C. for at least 30 minutes or until a slight yellow coloration appeared. The reaction was stopped by adding 100 μl of 1 M Na₂CO₃, and the absorption was determined at 420 nm.

In order to analyse the hTC promoter, four hTC promoter sequence segments of differing length were cloned 5′ in front of the luciferase reporter gene (cf. Example 9).

The relative luciferase activities of two independent transfections in HEK 293 cells, using the constructs NPK8, NPK15, NPK22 and NPK27, are plotted in FIG. 11. Each experiment was carried out in duplicate. The standard deviation has also been given. The construct NPK 27 exhibits a luciferase activity which is 40 times higher than the basal activity of the promoterless luciferase control construct (pGL2-basic) and from 2 to 3 times higher than that of the SV40 promoter control construct (pGL2PRO). Interestingly, a luciferase activity which was from 2 to 3 times lower than that obtained with the NPK 27 construct was observed in cells which were transfected with longer hTC promoter constructs (NPK8, NPK15, NPK22). Similar results were also observed in CHO cells (data not shown).

REFERENCES

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1. Regulatory DNA sequences for the gene for the human catalytic telomerase subunit.
 2. DNA sequences according to claim 1, characterized in that the sequences are intron sequences in accordance with SEQ ID NO 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and/or 20 or fragments of these sequences which have a regulatory effect.
 3. DNA sequences according to claim 1, characterized in that the sequences are the 5′-flanking regulatory DNA sequence for the gene for the human catalytic telomerase subunit as depicted in FIG. 10 (SEQ ID NO 3), or fragments of this DNA sequence which have a regulatory effect.
 4. Recombinant construct which contains a DNA. sequence according to one of claims 1 to
 3. 5. Recombinant construct according to claim 4, characterized in that it additionally contains one or more DNA sequences which encode polypeptides or proteins.
 6. Vector which contains a recombinant construct according to claim 4 or
 5. 7. Use of recombinant constructs or vectors according to one of claims 4 to 6 for preparing medicaments.
 8. Recombinant host cells which harbour recombinant constructs or vectors according to one of claims 4 to
 6. 9. Process for identifying substances which affect the promoter activity, silencer activity or enhancer activity of the human catalytic telomerase subunit, comprising the following steps: A. adding a candidate substance to a host cell which harbours DNA sequences according to one of claims 1 to 3, which sequences are functionally linked to a reporter gene, and B. measuring the effect of the substance on expression of the reporter gene.
 10. Process for identifying factors which bind specifically to the DNA according to one of claims 1 to 3, or to fragments thereof, characterized in that an expression cDNA library is screened using a DNA sequence according to one of claims 1 to 3, or subfragments of widely differing length, as the probe.
 11. Transgenic animals which harbour recombinant constructs or vectors according to claims 4 to
 6. 12. Process for detecting telomerase-associated conditions in a patient, comprising the following steps: A. incubating a recombinant construct or vector according to claims 4 to 6, which additionally contains a reporter gene, with body fluids or cell samples, B. detecting the activity of the reporter gene in order to obtain a diagnostic value, and C. comparing the diagnostic value with standard values for the reporter gene construct in standardized normal cells or body fluids of the same type as the test sample. 